Research Article Identification and Validation of ...downloads.hindawi.com/archive/2014/651912.pdfInternationalJournalofPlantGenomics T :SummaryofESTslibrary. Total clone sequenced

Research ArticleIdentification and Validation of Expressed Sequence Tags fromPigeonpea (Cajanus cajan L) Root

Ravi Ranjan Kumar12 Shailesh Yadav3 Shourabh Joshi3 Prithviraj P Bhandare2

Vinod Kumar Patil2 Pramod B Kulkarni2 Swati Sonkawade2 and G R Naik2

1 Vidya Pratishthanrsquos School of Biotechnology Vidyanagari Baramati Pune 413133 India2Department of Biotechnology Gulbarga University Gulbarga Karnataka 585106 India3 Institute of Biotechnology Acharya N G Ranga Agricultural University Hyderabad 500030 India

Correspondence should be addressed to G R Naik grnaikbiotechgmailcom

Received 14 November 2013 Accepted 15 April 2014 Published 6 May 2014

Academic Editor Soslashren K Rasmussen

Copyright copy 2014 Ravi Ranjan Kumar et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Pigeonpea (Cajanus cajan (L) Millsp) is an important food legume crop of rain fed agriculture in the arid and semiarid tropicsof the world It has deep and extensive root system which serves a number of important physiological and metabolic functions inplant development and growth In order to identify genes associated with pigeonpea root ESTs were generated from the root tissuesof pigeonpea (GRG-295 genotype) by normalized cDNA library A total of 105 high quality ESTs were generated by sequencing of250 random clones which resulted in 72 unigenes comprising 25 contigs and 47 singlets The ESTs were assigned to 9 functionalcategories on the basis of their putative function In order to validate the possible expression of transcripts four genes namelyS-adenosylmethionine synthetase phosphoglycerate kinase serine carboxypeptidase and methionine aminopeptidase were furtheranalyzed by reverse transcriptase PCR The possible role of the identified transcripts and their functions associated with root willalso be a valuable resource for the functional genomics study in legume crop

1 Introduction

Pigeonpea (Cajanus cajan L) Millsp (2119899 = 22) is a majorgrain legume of the arid and semiarid regions of the world [1]Though considered a minor crop pigeonpea is of consider-able importance in areas of South Asia (mainly on the Indiansubcontinent) Africa the Caribbean and Latin Americawhere it is a prominent source of protein in the human dietas well as wood for fuel and light duty structural applicationssuch as thatch for roofing [2] Pigeonpea has nowmoved froman ldquoorphan legume croprdquo to one of the promising pluseswheregenomics-assisted breeding approaches for a sustainable cropimprovement are routine by Pigeonpea Genome Initiativean effort of various researchers [3] The first pigeonpea ESTdataset provides a transcriptomic resource for gene discoveryand development of functionalmarkers associatedwith bioticstress resistance [4] Root is the major part of water andnutrition uptake in pigeonpea which has a deep and extensiveroot system that provides access to water stored deep in

the soil profile when that in the surface layer is depleted thissource of water is particularly important for long durationcrops In order to identify the associated genes in pigeonpearoot tissues a normalized cDNA library was constructedfrom pigeonpea root and expression analysis of the identifiedgenes was carried out by reverse transcriptase PCR (RT-PCR)technique

2 Materials and Methods

The pigeonpea genotype namely GRG-295 was selectedto construct cDNA library and identification of expressedsequence tags (ESTs) The seeds of GRG-295 pigeonpeagenotype were grown in petri dish for 15 days and irrigatedwith water At the end of the 15th day total RNA wasisolated from root tissues by using the Trizol reagent(Invitrogen Carlsbad CA USA) and mRNA was furtherisolated by using the PolyATract mRNA Isolation System

Hindawi Publishing CorporationInternational Journal of Plant GenomicsVolume 2014 Article ID 651912 7 pageshttpdxdoiorg1011552014651912

2 International Journal of Plant Genomics

Table 1 Primer sequences of subset of ESTs for RT-PCR analysis

Accession number Putative function Optimum 119879119898(∘C) Primer sequence (51015840-31015840)

JK973671 S-Adenosylmethionine synthetase 61 F-AGAGGAAAT CGGT GCTGGTGR-GCAGCAATTTGGTCGTTGGT

JK973674 Phosphoglycerate kinase 59 F-TCCCGATCCCGATACCCTACR-CAGCACGCTTTTCAGCAGTT

JK973715 Serine carboxypeptidase 60 F-ACATGAAGCTCAGTGGAGGAGR-AGCCATGGCCTCCAATCTTC

JK973726 Methionine aminopeptidase 60 F-GGCAT TGAAAGTTGGGCAGGR-GATTGCAGCACCGACATCAC

(Promega Madison WI USA) The quantification of RNAwas verified by absorption ratio of OD

260280and by for-

maldehyde gel electrophoresis The first and second strandsof cDNA were synthesized using Clontech SMARTer PCRcDNA Synthesis Kit The cDNAs were purified by the Min-Elute PCR purification kit (Qiagen Valencia CA USA) andligated into a pGEM-T easy vector (Promega Madison WIUSA) Ligated plasmid DNAs were used for transformationinto competent E coli DH5120572 strain Positive clones wereselected on an ampicillinIPTGX-Gal LB plate PlasmidDNA from positive clones were isolated by using REAL96 plasmid isolation kit (Qiagen Netherlands) andpurified DNA was used for single-pass Sanger sequencingby using M13FR universal sequencing primers on ABIsequencing machine 3500XL Genetic analyzer All the ESTswere processed using VecScreen (httpwwwncbinlmnihgovVecScreenVecScreenhtml) to remove vector and clon-ing oligo sequences and various contaminants to trim a highquality region Based on the qualified sequences the pre-dicted amino acid sequences were used to search for similarpeptide sequences to search for similar protein sequences inpublic database NCBI (httpwwwncbinlmnihgov) usingthe BLASTx search algorithm [5] by using default parametersof the program The similarity scores between the cDNAclones and known sequences were represented by BLASTxprobability E values Further the ESTs were classified intodifferent functional categories based on the knowledgeof biochemistry plant physiology and molecular biology(httpwwwMetaCycorg) GO (httpwwwebiacuk)and COG (httpwwwncbinlmnihgovCOG) tools andby searching related abstracts in PubMed

21 RT-PCR Analysis Total root RNAs isolated from pigeon-pea root tissues were used for reverse transcription poly-merase chain reaction (RT-PCR) analysis Genomic DNAcontamination was removed by DNase I First-strand cDNAwas synthesized from each 2120583g of total RNA sample usingClontech SMARTer PCR cDNA Synthesis Kit according tothe manufacturerrsquos protocol The cDNAs were purified usinga commercial column (Qiagen) To determine the expressionof candidate genes PCR was performed with 2120583L of thefirst-strand cDNA template and gene-specific primer pairsGene-specific RT-PCR primers were designed with Primer30 according to the EST sequences and were synthesizedcommercially General PCR was conducted with annealing

as required for the specific primer pairs (Table 1) RT-PCRexperimentswere repeated three times and the PCRproductswere detected on 15 agarose gel

3 Results and Discussion

Plant root systems serve a number of important functionsincluding anchoring the plant absorbingwater and nutrientsproducing amino acids and hormones and secreting organicacids enzymes and alkaloids [6] The physiological signifi-cance of roots is belied by their relative structural simplicityas compared to other plant organs major metabolic path-ways such as photosynthesis lacking in root tissues have astereotypical morphology that is conserved across taxa andthroughout the life cycle of individuals This combination ofphysiological relevance and structural simplicity has maderoots obvious targets for functional genomics analyses [7]As a major grain legume of semiarid tropics and a deep andextensive root system of pigeonpea represents an excellentsource of identification of ESTs associated with their roottissues So the presentworkwas focused on the study of genesassociated with pigeonpea root tissues

In the present investigation the cDNA library was con-structed in order to identify ESTs associated with pigeonpearoots and their functional analysis was carried out Thetotal RNA from the pigeonpea root tissue was isolatedand the first and second cDNA strand was synthesizedThe presence of the gene in plasmid construct of colonieswas confirmed by colony PCR The colony PCR showedthat the size of these inserts ranged from 200 to 800 bpOut of 400 bacterial clones the plasmid construct of 250positive recombinant clones was sequenced in single passedsequencing reaction from 31015840 end using M13 forwardreverseprimer and the sequence data was subjected to BLASTanalysis The leading sequences tailing of the sequence andpoor quality sequences were excluded firstly Finally 105 highquality ESTs were retained which were clustered into 72unigenes comprising 25 contigs and 47 singlets (Table 2) andwere compared with NCBI nonredundant protein databaseusing BLASTx algorithm and default parameters In BLASTxanalysis it was shown that most of the sequences were havinga significant homology with known proteins Sequences thathad no significant homology with protein database werecompared to nucleotide BLAST using default parameters

International Journal of Plant Genomics 3

Table 2 Summary of ESTs library

Total clone sequenced 250ESTs taken for analysis 105Number of unigenes 72Number of contigs 25Number of singlets 47Average length of unigenes 442 bpAverage length of ESTs 437 bp GC content of unigenes 503 GC content of ESTs 512

The ESTs were deposited to NCBI dbEST with the accessionnumber of JK973637 to JK973741 (Table 3)

TheESTswere categorized into 8 diverse functional groupconsisting of 10 transporter genes 6 signal transductiongenes 20 cell growth and transcriptional regulator genesand 26 metabolism genes In other genes 8 genes wereuncharacterized 8 hypothetical genes 10 genes with nosignificant match and 14 genes was involved in otherfunctionsThere were 8 of the ESTs found that did not showany match to known proteins in BLASTx program and thatsuggest novel nature of those genes (Figure 1)

In order to validate the ESTs generated from cDNAlibrary the expression of the four genes which were involvedin different metabolic pathways was analysed by RT-PCRRT-PCR results showed that the expression levels of fourcandidate genes namely S-adenosylmethionine synthetasephosphoglycerate kinase serine carboxypeptidase andmethio-nine aminopeptidase were clearly expressed in pigeonpearoots (Figure 2) It was concluded that overall there was agood agreement between the cDNA library data and the RT-PCR results

S-Adenosylmethionine synthetase (SAMS) comprises oftwo cDNAs in Pinus contorta among which SAMS1 isexpressed in roots and exhibits a specific expression patternin the meristem at the onset of adventitious root devel-opment [8] SAMS also catalyses the nucleophilic substi-tution reaction from between methionine and ATP intoS-adenosylmethionine which have central role in severalbiological process in plants namely methyl group donor intrimethylation of lignin DNA and alkaloids as well as donorof aminopropylmoieties in ethylene and polyamine synthesis[8 9]

Phosphoglycerate kinase superfamily has a diverse func-tion in numerous metabolic processes like generation ofprecursor metabolites and energy carbohydrate metabolismphosphorous metabolism glycolysis kinase activity ATPbinding activity and so forth The presence of phosphoglycer-ate kinase transcripts in the cDNA library supports its diversefunction in pigeonpea root

Serine carboxypeptidase differentially expressed in rootand other tissues is responsible for the synthesis of sinapoyl-choline and sinapoylmalate in Arabidopsis which encodes 51proteins annotated as serine carboxypeptidase-like enzymesand emerged as a new group of acyltransferase enzymesthat are able to modify plant natural products [10ndash12]

Hyp protein8

Transpoter10

Signaltransduction

6

Cell wall synthesis and transcriptional

regulator20

Metabolism26

Uncharacterized protein

8

Other functions14

No hits8

Figure 1 Functional classification of ESTs of pigeonpea generatedby single pass sequencing

M 4321

Figure 2 Validation of cDNA library results by RT-PCR (M-100 bp ladder 1-S-adenosylmethionine synthetase 2-phosphoglyceratekinase 3-serine carboxypeptidase and 4-methionine aminopepti-dase)

Methionine aminopeptidase a ubiquitous enzyme differen-tially expressed in root tissues is one of the central enzymes inprotein synthesis that catalyzes N-terminal methionine fromproteins [13]

A few important ESTs viz type 2 metallothionein TBP-associated factor ABC transporter and phosphatidylcholinetransfer protein were also abundantly found in the cDNAlibrary Reddy et al [14] found abundant metallothioneingenes in normalized library from rice leaves and suggestedthat they might perform essential functions of plant growthbeside metal detoxification TATA-binding protein and TBP-associated factors are transcriptional factors which are pre-dominantly involved in RNA polymerase II mediated tran-scription process [15] ABC transporters play an importantrole in organ growth plant nutrition plant developmentresponse to abiotic stress and the interaction of the plantwith its environment [16] Phosphatidylcholine is usuallythe most abundant phospholipids in animals and plantsoften amounting to almost 50 of the total and as such itis obviously the key building block of membrane bilayersmaking up a very high proportion of the outer leaflet of


Table3Hom

olog

ysearch

oftheE

STsg

enerated

from

pigeon

pear

ootalong

with

their119864

valueu

singBL

AST

xprogramme

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

1JK

973637

560

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

2JK

973638

231

Mito

chon

drialinn

ermem

branep

rotein

OXA1-like[

Vitis

vinifer

a]5119890minus41

3JK

973639

630

Uncharacterized

protein[Arabidopsisthaliana

]9119890minus159

4JK

973640

621

Hypothetic

alprotein[Sorghum

bicolor]

9119890minus139

5JK

973641

570

GTP

-binding

proteinhfl

x[M

edica

gotru

ncatula]

6119890minus13

6JK

973642

412

Unk

nown[G

lycinem

ax]

11

7JK

973643

264

Secretorycarrier-associated

mem

branep

rotein

3-lik

e[Vitis

vinifer

a]2119890minus55

8JK

973644

438

Histon

e2[Populus

trichocarpa]

5119890minus63

9JK

973645

200

RNArecogn

ition

motif-containing


]4119890minus10

10JK973646

870

Beta-glucosid

ase[

Arabidopsis

thaliana

]00

11JK

973647

555

Epsin

N-te

rminalho

molog

y(ENTH

)dom

ain-containing

protein[M

edica

gotru

ncatula]

4119890minus36

12JK

973648

210

Gam

ma-gliadinprecursor[Ricin

uscommun

is]7119890minus10

13JK

973649

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor

[Arabidopsisthaliana

]4119890minus40

14JK

973650

474

Glycine

dehydrogenase(decarborcylatingmito

chon

driallike)[G

lycinem

ax]

4119890minus102

15JK

973651

432

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

16JK

973652

227

TBP-associated

factor

6B[Arabidopsisthaliana

]0051

17JK

973653

290

TBP-associated

factor


]8119890minus48

18JK

973654

303

Transcrip

tionfactor

[Medica

gotru

ncatula]

6119890minus29

19JK

973655

432

Uncharacterized

protein[G

lycinem

ax]

8119890minus11

20JK

973656

237

Nuclear

cap-bind

ingproteinsubu

nit[Arabidopsis

thaliana

]8119890minus48

21JK

973657

501

Glucanendo

-13-beta-glucosidase-lik

eprotein

3-lik

e[Glycinem

ax]

2119890minus04

22JK

973658

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

23JK

973659

560

Two-compo

nent

respon

seregulator

ARR

9[Vitisv

inifera]

1119890minus45

24JK

973660

210

Ribo

nucle

oside-diph

osph

ater

eductase

smallchain

like[Glycinem

ax]

2119890minus34

25JK

973661

748

Hypothetic

alprotein[O

ryza

sativa]

26JK

973662

742

Cyclin-depend

entp

rotein

kinase

complex

compo

nent

[Aspergillu

skaw

achii]

012

27JK

973663

406

Elon

gatio

nfactor

2[N

icotia

natobacum]

5119890minus39

28JK

973664

841

Expressedprotein[O

ryza

sativa]

73

29JK

973665

385

Hypothetic

alprotein[Vitisv

inifera]

21

30JK

973666

544

ADP-rib

osylationfactor-like

8d[N

icotia

natobacum]

2119890minus105

31JK

973667

510

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

6119890minus77

32JK

973668

630

Nosignific

antm

atch

33JK

973669

804

Gag-pro[Pisu

msativ

um]

9119890minus18

34JK

973670

320

HypotheticalproteinMTR

[Medica

gotru

ncatula]

2119890minus13

35JK

973671

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

m]

6119890minus114

36JK

973672

371

ABC

transporterF

family

mem

ber1-like

[Brachypodium

dista

chyon]

4119890minus60

37JK

973673

513

Predictedprotein[H

ordeum

vulga

re]

2119890minus89

38JK973674

465

Phosph

oglyceratekinasecytosolic[Triticu

maestivum]

2119890minus97

39JK973675

465

Phosph


maestivum]

2119890minus97

40JK

973676

457

Unk

nown[Pice

asitchensis]

097

41JK

973677

785

GTP

-binding

signalrecognitio

nparticleSR

P54[M

edica

gotru

ncatula]

7119890minus51


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

42JK

973678

241

Sign

alrecogn

ition

particle54

kDAsubu

nitp

recursor

[Pisu

msativ

um]

0044

43JK

973679

475

ProteinSE

T-lik

e[Brachypodium

dista

chyon]

7119890minus33

44JK

973680

350

EIN3-bind

ingF-bo

xprotein[Brachypodium

dista

chyon]

8119890minus65

45JK

973681

389

Predictedprotein[H

ordeum

vulga

re]

4119890minus27

46JK

973682

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

47JK973683

210

Putativ

ecalcium

exchanger[Triticum

dicoccoides]

4119890minus27

48JK

973684

478

Thioredo

xin[M

edica

gotru

ncatula]

2119890minus75

49JK

973685

210

Nosignific

antm

atch

50JK

973686

553

Root

nodu

leextension[Pisu

msativa]

5119890minus04

51JK

973687

600

Sign

alrecogn

ition

particleprotein[O

ryza

sativa]

5119890minus08

52JK

973688

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

53JK

973689

395

MLO

-like

protein[M

edica

gotru

ncatula]

2119890minus24

54JK973690

250

Phosph

atidylcholinetransferp

rotein

[Ricinu

scom

mun

is]3119890minus09

55JK973691

215

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

56JK

973692

411

Unk

nown[G

lycinem

ax]

11

57JK973693

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

58JK973694

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

59JK973695

420

Uncharacterized

protein[Zea

mays]

9119890minus83

60JK

973696

350

Uncharacterized

protein[Zea

mays]

5119890minus51

61JK

973697

244

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

5119890minus27

62JK

973698

415

Os01g0678900[O

ryza

sativa]

8119890minus11

63JK

973699

282

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]3119890minus51

64JK

973700

558

60Srib

osom

alprotein[Vitisv

inifera]

8119890minus65

65JK

973701

490

Nosignific

antm

atch

66JK

973702

556

Root

nodu

leextension[Pisu

msativ

um]

3119890minus07

67JK

973703

313

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]2119890minus65

68JK

973704

311

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]1119890minus64

69JK

973705

320

Hypothetic

alproteinMTR

4g076190

[Medica

gotru

ncatula]

2119890minus13

70JK

973706

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

um]

9119890minus119

71JK

973707

460

Transm

embranee

mp24do

main-containing

protein10-like

[Brachypodium

dista

chyon]

2119890minus45

72JK

973708

482

60Srib

osom

alprotein[Zea

mays]

3119890minus52

73JK

973709

395

MLO

5-lik

eprotein

[Medica

gotru

ncatula]

2119890minus24

74JK

973710

480

Type

2metallothionein

[Prosopisjulifl

ora]

4119890minus28

75JK

973711

556

Root

nodu

leextension[Pisu

msativa]

3119890minus07

76JK

973712

174

Hypothetic

alprotein[O

ryza

sativa]

086

77JK

973713

278

Nosignific

antm

atch

78JK

973714

439

Nosignific

antm

atch

79JK

973715

439

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

2119890minus31

80JK

973716

551

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

9119890minus39

81JK

973717

347

Nosignific

antm

atch

82JK

973718

661

Gam

ma-glutam

ylhydrolase[Vitis

vinifer

a]1119890minus54

83JK

973719

499

Type

2metallothionein

[Prosopisjuliflora]

2119890minus30


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

84JK

973720

463

Nod

ulin

mtn21eam

a-lik

etranspo

rter


]2119890minus44

85JK

973721

200

RNArecogn

ition

motif-containing


]4119890minus10

86JK

973722

280

Beta-glucosid

ase4

4-lik

e[Glycinem

ax]

3119890minus52

87JK

973723

424

V-type

proton

ATpase

21kD

Aproteolip

idsubu

nit[Medica

gotru

ncatula]

2119890minus29

88JK

973724

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

89JK

973725

540

Polyrib

onucleotiden

ucleotidyltransfe

rase

[Vitisv

inifera]

3119890minus07

90JK

973726

649

Methion

inea

minop

eptid

ase2

B-lik

e[Brachypodium

dista

chyon]

3119890minus40

91JK

973727

585

Hypothetic

alprotein[Vitisvinifera]

69

92JK

973728

487

Unk

nown[Lotus

japonica]

1

93JK

973729

558

60Srib

osom


8119890minus65

94JK

973730

314

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]1119890minus55

95JK

973731

350

EIN3-bind

ingF-bo

xprotein1-like[Brachypodium

dista

chyon]

8119890minus65

96JK

973732

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

97JK

973733

239

NOT2

NOT3

NOT5

family

protein[O

ryza

sativa]

3119890minus62

98JK

973734

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

99JK

973735

559

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

100

JK973736

793

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

101

JK973737

649

Methion

inea

minop

eptid

ase2

B[Brachypodium

dista

chyon]

1119890minus24

102

JK973738

210

Gam


uscommun

is]7119890minus10

103

JK973739

553

Unk

nownproteinprod

uct[Glycinem

ax]

1119890minus04

104

JK973740

600

Glycine

dehydrogenase[Glycinem

ax]

4119890minus102

105

JK973741

572

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04


the plasma membrane [17] Similarly numerous ESTs likeglycine dehydrogenase signal proteins root nodule exten-sions membrane proteins and 120573-glucosidase were observedin the cDNA library constructed from pigeonpea root tissueswhich is thought to play possible roles in plant metabolismgrowth and development

Apart from the knownESTs 6 EST transcripts (JK973668JK973685 JK973701 JK973714 JK973715 and JK973718)wereobserved as not having any significant match in NCBI data-base Along with that uncharacterized and hypothetical pro-teins were also observed in cDNA library which were thoughtto impart in the cardinal role in root tissues of pigeonpea

4 Conclusion

Our present investigation contains a precise repertoire oftranscripts associated with the various metabolic functionsin pigeonpea root These ESTs appear to be involved in mul-tiple metabolism pathways in the plantrsquos physiological andbiochemical processes In addition to known genes someESTs were unknown and uncharacterized whose functionalroles remain unclear and require further investigation infuture The root transcriptome characterized in this studymarkedly provides a unique resource for investigating thefunctional specificities of the root systemThese EST tagsmaybe useful for functional gene annotation analysis of splicesite variants and intronexon determination and evaluationof gene homologies or KEGG pathway confirmation

Abbreviations

ESTs Expressed sequence tagsRT PCR Reverse transcription polymerase chain

reactionSAMS S-Adenosyl methionine synthetase

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] Y L Nene and V K Shiela ldquoPigeon pea geography and impor-tancerdquo inThePigeon Pea Y L Nene S H Hall and V K SheilaEds pp 1ndash14 CAB International Wellingford UK 1990

[2] M T Kassa R V Penmetsa N Carrasquilla-Garcia et alldquoGenetic patterns of domestication in pigeonpea (Cajanus cajan(L) Millsp) and wild Cajanus relativesrdquo PLoS ONE vol 7 no6 Article ID e39563 2012

[3] R K Varshney R V Penmetsa S Dutta et al ldquoPigeonpeagenomics initiative (PGI) an international effort to improvecrop productivity of pigeonpea (Cajanus cajan L)rdquo MolecularBreeding vol 26 no 3 pp 393ndash408 2010

[4] N L Raju B N Gnanesh P Lekha et al ldquoThe first set of ESTresource for gene discovery andmarker development in pigeon-pea (Cajanus cajan L)rdquo BMC Plant Biology vol 10 pp 45ndash512010

[5] S F Altschul T L Madden A A Schaffer et al ldquoGappedBLAST and PSI-BLAST a new generation of protein databasesearch programsrdquo Nucleic Acids Research vol 25 no 17 pp3389ndash3402 1997

[6] R Zhai Y Feng H Wang et al ldquoTranscriptome analysis of riceroot heterosis by RNA-Seqrdquo BMC Genomics vol 14 article 19pp 1ndash14 2013

[7] Y Jiang and M K Deyholos ldquoComprehensive transcriptionalprofiling of NaCl-stressed Arabidopsis roots reveals novelclasses of responsive genesrdquo BMC Plant Biology vol 6 article25 2006

[8] A M Lindroth P Saarikoski G Flygh et al ldquoTwo S-adenosyl-methionine synthetase-encoding genes differentially expressedduring adventitious root development in Pinus contortardquo PlantMolecular Biology vol 46 no 3 pp 335ndash346 2001

[9] P K Chiang R K Gordon J Tal et al ldquoS-adenosylmethionineandmethylationrdquoTheFASEB Journal vol 10 no 4 pp 471ndash4801996

[10] A M Shirley C M McMichael and C Chapple ldquoThe sng2mutant of Arabidopsis is defective in the gene encoding theserine carboxypeptidase-like protein sinapoylglucosecholinesinapoyltransferaserdquo Plant Journal vol 28 no 1 pp 83ndash942001

[11] C M Fraser M G Thompson A M Shirley et al ldquoRelatedArabidopsis serine carboxypeptidase-like sinapoylglucose acyl-transferases display distinct but overlapping substrate specifici-tiesrdquo Plant Physiology vol 144 no 4 pp 1986ndash1999 2007

[12] D Weier J Mittasch D Strack and C Milkowski ldquoThe genesBnSCT1 and BnSCT2 from Brassica napus encoding the finalenzyme of sinapine biosynthesis molecular characterizationand suppressionrdquo Planta vol 227 no 2 pp 375ndash385 2008

[13] W T Lowther and B W Matthews ldquoStructure and functionof the methionine aminopeptidasesrdquo Biochimica et BiophysicaActa vol 1477 no 1-2 pp 157ndash167 2000

[14] V S Reddy G S Ali and A S N Reddy ldquoGenes encodingcalmodulin-binding proteins in the Arabidopsis genomerdquo Jour-nal of Biological Chemistry vol 277 no 12 pp 9840ndash9852 2002

[15] S R Albright and R Tjian ldquoTAFs revisited more data revealnew twists and confirm old ideasrdquo Gene vol 242 no 1-2 pp1ndash13 2000

[16] J Kang J Park H Choi et al ldquoPlant ABC transportersrdquo TheArabidopsis Book vol 9 pp 1ndash25 2010

[17] K A Devor and J B Mudd ldquoStructural analysis of phosphat-idylcholine of plant tissuerdquo Journal of Lipid Research vol 12 no4 pp 396ndash402 1971

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


Table 1 Primer sequences of subset of ESTs for RT-PCR analysis

Accession number Putative function Optimum 119879119898(∘C) Primer sequence (51015840-31015840)

JK973671 S-Adenosylmethionine synthetase 61 F-AGAGGAAAT CGGT GCTGGTGR-GCAGCAATTTGGTCGTTGGT

JK973674 Phosphoglycerate kinase 59 F-TCCCGATCCCGATACCCTACR-CAGCACGCTTTTCAGCAGTT

JK973715 Serine carboxypeptidase 60 F-ACATGAAGCTCAGTGGAGGAGR-AGCCATGGCCTCCAATCTTC

JK973726 Methionine aminopeptidase 60 F-GGCAT TGAAAGTTGGGCAGGR-GATTGCAGCACCGACATCAC

(Promega Madison WI USA) The quantification of RNAwas verified by absorption ratio of OD

260280and by for-

maldehyde gel electrophoresis The first and second strandsof cDNA were synthesized using Clontech SMARTer PCRcDNA Synthesis Kit The cDNAs were purified by the Min-Elute PCR purification kit (Qiagen Valencia CA USA) andligated into a pGEM-T easy vector (Promega Madison WIUSA) Ligated plasmid DNAs were used for transformationinto competent E coli DH5120572 strain Positive clones wereselected on an ampicillinIPTGX-Gal LB plate PlasmidDNA from positive clones were isolated by using REAL96 plasmid isolation kit (Qiagen Netherlands) andpurified DNA was used for single-pass Sanger sequencingby using M13FR universal sequencing primers on ABIsequencing machine 3500XL Genetic analyzer All the ESTswere processed using VecScreen (httpwwwncbinlmnihgovVecScreenVecScreenhtml) to remove vector and clon-ing oligo sequences and various contaminants to trim a highquality region Based on the qualified sequences the pre-dicted amino acid sequences were used to search for similarpeptide sequences to search for similar protein sequences inpublic database NCBI (httpwwwncbinlmnihgov) usingthe BLASTx search algorithm [5] by using default parametersof the program The similarity scores between the cDNAclones and known sequences were represented by BLASTxprobability E values Further the ESTs were classified intodifferent functional categories based on the knowledgeof biochemistry plant physiology and molecular biology(httpwwwMetaCycorg) GO (httpwwwebiacuk)and COG (httpwwwncbinlmnihgovCOG) tools andby searching related abstracts in PubMed

21 RT-PCR Analysis Total root RNAs isolated from pigeon-pea root tissues were used for reverse transcription poly-merase chain reaction (RT-PCR) analysis Genomic DNAcontamination was removed by DNase I First-strand cDNAwas synthesized from each 2120583g of total RNA sample usingClontech SMARTer PCR cDNA Synthesis Kit according tothe manufacturerrsquos protocol The cDNAs were purified usinga commercial column (Qiagen) To determine the expressionof candidate genes PCR was performed with 2120583L of thefirst-strand cDNA template and gene-specific primer pairsGene-specific RT-PCR primers were designed with Primer30 according to the EST sequences and were synthesizedcommercially General PCR was conducted with annealing

as required for the specific primer pairs (Table 1) RT-PCRexperimentswere repeated three times and the PCRproductswere detected on 15 agarose gel

3 Results and Discussion

Plant root systems serve a number of important functionsincluding anchoring the plant absorbingwater and nutrientsproducing amino acids and hormones and secreting organicacids enzymes and alkaloids [6] The physiological signifi-cance of roots is belied by their relative structural simplicityas compared to other plant organs major metabolic path-ways such as photosynthesis lacking in root tissues have astereotypical morphology that is conserved across taxa andthroughout the life cycle of individuals This combination ofphysiological relevance and structural simplicity has maderoots obvious targets for functional genomics analyses [7]As a major grain legume of semiarid tropics and a deep andextensive root system of pigeonpea represents an excellentsource of identification of ESTs associated with their roottissues So the presentworkwas focused on the study of genesassociated with pigeonpea root tissues

In the present investigation the cDNA library was con-structed in order to identify ESTs associated with pigeonpearoots and their functional analysis was carried out Thetotal RNA from the pigeonpea root tissue was isolatedand the first and second cDNA strand was synthesizedThe presence of the gene in plasmid construct of colonieswas confirmed by colony PCR The colony PCR showedthat the size of these inserts ranged from 200 to 800 bpOut of 400 bacterial clones the plasmid construct of 250positive recombinant clones was sequenced in single passedsequencing reaction from 31015840 end using M13 forwardreverseprimer and the sequence data was subjected to BLASTanalysis The leading sequences tailing of the sequence andpoor quality sequences were excluded firstly Finally 105 highquality ESTs were retained which were clustered into 72unigenes comprising 25 contigs and 47 singlets (Table 2) andwere compared with NCBI nonredundant protein databaseusing BLASTx algorithm and default parameters In BLASTxanalysis it was shown that most of the sequences were havinga significant homology with known proteins Sequences thathad no significant homology with protein database werecompared to nucleotide BLAST using default parameters










Hyp protein8

Transpoter10

Signaltransduction

6


regulator20

Metabolism26


8

Other functions14

No hits8


M 4321





Table3Hom

olog

ysearch

oftheE

STsg

enerated

from

pigeon

pear

ootalong

with

their119864

valueu

singBL

AST

xprogramme

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

1JK

973637

560

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

2JK

973638

231

Mito

chon

drialinn

ermem

branep

rotein

OXA1-like[

Vitis

vinifer

a]5119890minus41

3JK

973639

630

Uncharacterized


]9119890minus159

4JK

973640

621

Hypothetic

alprotein[Sorghum

bicolor]

9119890minus139

5JK

973641

570

GTP

-binding

proteinhfl

x[M

edica

gotru

ncatula]

6119890minus13

6JK

973642

412

Unk

nown[G

lycinem

ax]

11

7JK

973643

264


mem

branep

rotein

3-lik

e[Vitis

vinifer

a]2119890minus55

8JK

973644

438

Histon

e2[Populus

trichocarpa]

5119890minus63

9JK

973645

200

RNArecogn

ition

motif-containing


]4119890minus10

10JK973646

870

Beta-glucosid

ase[

Arabidopsis

thaliana

]00

11JK

973647

555

Epsin

N-te

rminalho

molog

y(ENTH

)dom

ain-containing

protein[M

edica

gotru

ncatula]

4119890minus36

12JK

973648

210

Gam


uscommun

is]7119890minus10

13JK

973649

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

14JK

973650

474

Glycine


chon

driallike)[G

lycinem

ax]

4119890minus102

15JK

973651

432

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

16JK

973652

227

TBP-associated

factor


]0051

17JK

973653

290

TBP-associated

factor


]8119890minus48

18JK

973654

303

Transcrip

tionfactor

[Medica

gotru

ncatula]

6119890minus29

19JK

973655

432

Uncharacterized

protein[G

lycinem

ax]

8119890minus11

20JK

973656

237

Nuclear

cap-bind

ingproteinsubu

nit[Arabidopsis

thaliana

]8119890minus48

21JK

973657

501

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04

22JK

973658

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

23JK

973659

560

Two-compo

nent

respon

seregulator

ARR

9[Vitisv

inifera]

1119890minus45

24JK

973660

210

Ribo

nucle

oside-diph

osph

ater

eductase

smallchain

like[Glycinem

ax]

2119890minus34

25JK

973661

748

Hypothetic

alprotein[O

ryza

sativa]

26JK

973662

742

Cyclin-depend

entp

rotein

kinase

complex

compo

nent

[Aspergillu

skaw

achii]

012

27JK

973663

406

Elon

gatio

nfactor

2[N

icotia

natobacum]

5119890minus39

28JK

973664

841

Expressedprotein[O

ryza

sativa]

73

29JK

973665

385

Hypothetic

alprotein[Vitisv

inifera]

21

30JK

973666

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

31JK

973667

510

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

6119890minus77

32JK

973668

630

Nosignific

antm

atch

33JK

973669

804

Gag-pro[Pisu

msativ

um]

9119890minus18

34JK

973670

320


[Medica

gotru

ncatula]

2119890minus13

35JK

973671

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

m]

6119890minus114

36JK

973672

371

ABC

transporterF

family

mem

ber1-like

[Brachypodium

dista

chyon]

4119890minus60

37JK

973673

513

Predictedprotein[H

ordeum

vulga

re]

2119890minus89

38JK973674

465

Phosph


maestivum]

2119890minus97

39JK973675

465

Phosph


maestivum]

2119890minus97

40JK

973676

457

Unk

nown[Pice

asitchensis]

097

41JK

973677

785

GTP

-binding

signalrecognitio

nparticleSR

P54[M

edica

gotru

ncatula]

7119890minus51


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

42JK

973678

241

Sign

alrecogn

ition

particle54

kDAsubu

nitp

recursor

[Pisu

msativ

um]

0044

43JK

973679

475

ProteinSE

T-lik

e[Brachypodium

dista

chyon]

7119890minus33

44JK

973680

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

45JK

973681

389

Predictedprotein[H

ordeum

vulga

re]

4119890minus27

46JK

973682

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

47JK973683

210

Putativ

ecalcium

exchanger[Triticum

dicoccoides]

4119890minus27

48JK

973684

478

Thioredo

xin[M

edica

gotru

ncatula]

2119890minus75

49JK

973685

210

Nosignific

antm

atch

50JK

973686

553

Root

nodu

leextension[Pisu

msativa]

5119890minus04

51JK

973687

600

Sign

alrecogn

ition

particleprotein[O

ryza

sativa]

5119890minus08

52JK

973688

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

53JK

973689

395

MLO

-like

protein[M

edica

gotru

ncatula]

2119890minus24

54JK973690

250

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

55JK973691

215

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

56JK

973692

411

Unk

nown[G

lycinem

ax]

11

57JK973693

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

58JK973694

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

59JK973695

420

Uncharacterized

protein[Zea

mays]

9119890minus83

60JK

973696

350

Uncharacterized

protein[Zea

mays]

5119890minus51

61JK

973697

244

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

5119890minus27

62JK

973698

415

Os01g0678900[O

ryza

sativa]

8119890minus11

63JK

973699

282

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]3119890minus51

64JK

973700

558

60Srib

osom

alprotein[Vitisv

inifera]

8119890minus65

65JK

973701

490

Nosignific

antm

atch

66JK

973702

556

Root

nodu

leextension[Pisu

msativ

um]

3119890minus07

67JK

973703

313

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]2119890minus65

68JK

973704

311

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]1119890minus64

69JK

973705

320

Hypothetic

alproteinMTR

4g076190

[Medica

gotru

ncatula]

2119890minus13

70JK

973706

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

um]

9119890minus119

71JK

973707

460

Transm

embranee

mp24do

main-containing

protein10-like

[Brachypodium

dista

chyon]

2119890minus45

72JK

973708

482

60Srib

osom

alprotein[Zea

mays]

3119890minus52

73JK

973709

395

MLO

5-lik

eprotein

[Medica

gotru

ncatula]

2119890minus24

74JK

973710

480

Type

2metallothionein

[Prosopisjulifl

ora]

4119890minus28

75JK

973711

556

Root

nodu

leextension[Pisu

msativa]

3119890minus07

76JK

973712

174

Hypothetic

alprotein[O

ryza

sativa]

086

77JK

973713

278

Nosignific

antm

atch

78JK

973714

439

Nosignific

antm

atch

79JK

973715

439

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

2119890minus31

80JK

973716

551

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

9119890minus39

81JK

973717

347

Nosignific

antm

atch

82JK

973718

661

Gam

ma-glutam

ylhydrolase[Vitis

vinifer

a]1119890minus54

83JK

973719

499

Type

2metallothionein

[Prosopisjuliflora]

2119890minus30


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

84JK

973720

463

Nod

ulin

mtn21eam

a-lik

etranspo

rter


]2119890minus44

85JK

973721

200

RNArecogn

ition

motif-containing


]4119890minus10

86JK

973722

280

Beta-glucosid

ase4

4-lik

e[Glycinem

ax]

3119890minus52

87JK

973723

424

V-type

proton

ATpase

21kD

Aproteolip

idsubu

nit[Medica

gotru

ncatula]

2119890minus29

88JK

973724

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

89JK

973725

540

Polyrib

onucleotiden

ucleotidyltransfe

rase

[Vitisv

inifera]

3119890minus07

90JK

973726

649

Methion

inea

minop

eptid

ase2

B-lik

e[Brachypodium

dista

chyon]

3119890minus40

91JK

973727

585

Hypothetic


69

92JK

973728

487

Unk

nown[Lotus

japonica]

1

93JK

973729

558

60Srib

osom


8119890minus65

94JK

973730

314

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]1119890minus55

95JK

973731

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

96JK

973732

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

97JK

973733

239

NOT2

NOT3

NOT5

family

protein[O

ryza

sativa]

3119890minus62

98JK

973734

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

99JK

973735

559

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

100

JK973736

793

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

101

JK973737

649

Methion

inea

minop

eptid

ase2

B[Brachypodium

dista

chyon]

1119890minus24

102

JK973738

210

Gam


uscommun

is]7119890minus10

103

JK973739

553

Unk

nownproteinprod

uct[Glycinem

ax]

1119890minus04

104

JK973740

600

Glycine


ax]

4119890minus102

105

JK973741

572

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04




4 Conclusion


Abbreviations





References

























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology










Hyp protein8

Transpoter10

Signaltransduction

6


regulator20

Metabolism26


8

Other functions14

No hits8


M 4321





Table3Hom

olog

ysearch

oftheE

STsg

enerated

from

pigeon

pear

ootalong

with

their119864

valueu

singBL

AST

xprogramme

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

1JK

973637

560

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

2JK

973638

231

Mito

chon

drialinn

ermem

branep

rotein

OXA1-like[

Vitis

vinifer

a]5119890minus41

3JK

973639

630

Uncharacterized


]9119890minus159

4JK

973640

621

Hypothetic

alprotein[Sorghum

bicolor]

9119890minus139

5JK

973641

570

GTP

-binding

proteinhfl

x[M

edica

gotru

ncatula]

6119890minus13

6JK

973642

412

Unk

nown[G

lycinem

ax]

11

7JK

973643

264


mem

branep

rotein

3-lik

e[Vitis

vinifer

a]2119890minus55

8JK

973644

438

Histon

e2[Populus

trichocarpa]

5119890minus63

9JK

973645

200

RNArecogn

ition

motif-containing


]4119890minus10

10JK973646

870

Beta-glucosid

ase[

Arabidopsis

thaliana

]00

11JK

973647

555

Epsin

N-te

rminalho

molog

y(ENTH

)dom

ain-containing

protein[M

edica

gotru

ncatula]

4119890minus36

12JK

973648

210

Gam


uscommun

is]7119890minus10

13JK

973649

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

14JK

973650

474

Glycine


chon

driallike)[G

lycinem

ax]

4119890minus102

15JK

973651

432

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

16JK

973652

227

TBP-associated

factor


]0051

17JK

973653

290

TBP-associated

factor


]8119890minus48

18JK

973654

303

Transcrip

tionfactor

[Medica

gotru

ncatula]

6119890minus29

19JK

973655

432

Uncharacterized

protein[G

lycinem

ax]

8119890minus11

20JK

973656

237

Nuclear

cap-bind

ingproteinsubu

nit[Arabidopsis

thaliana

]8119890minus48

21JK

973657

501

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04

22JK

973658

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

23JK

973659

560

Two-compo

nent

respon

seregulator

ARR

9[Vitisv

inifera]

1119890minus45

24JK

973660

210

Ribo

nucle

oside-diph

osph

ater

eductase

smallchain

like[Glycinem

ax]

2119890minus34

25JK

973661

748

Hypothetic

alprotein[O

ryza

sativa]

26JK

973662

742

Cyclin-depend

entp

rotein

kinase

complex

compo

nent

[Aspergillu

skaw

achii]

012

27JK

973663

406

Elon

gatio

nfactor

2[N

icotia

natobacum]

5119890minus39

28JK

973664

841

Expressedprotein[O

ryza

sativa]

73

29JK

973665

385

Hypothetic

alprotein[Vitisv

inifera]

21

30JK

973666

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

31JK

973667

510

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

6119890minus77

32JK

973668

630

Nosignific

antm

atch

33JK

973669

804

Gag-pro[Pisu

msativ

um]

9119890minus18

34JK

973670

320


[Medica

gotru

ncatula]

2119890minus13

35JK

973671

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

m]

6119890minus114

36JK

973672

371

ABC

transporterF

family

mem

ber1-like

[Brachypodium

dista

chyon]

4119890minus60

37JK

973673

513

Predictedprotein[H

ordeum

vulga

re]

2119890minus89

38JK973674

465

Phosph


maestivum]

2119890minus97

39JK973675

465

Phosph


maestivum]

2119890minus97

40JK

973676

457

Unk

nown[Pice

asitchensis]

097

41JK

973677

785

GTP

-binding

signalrecognitio

nparticleSR

P54[M

edica

gotru

ncatula]

7119890minus51


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

42JK

973678

241

Sign

alrecogn

ition

particle54

kDAsubu

nitp

recursor

[Pisu

msativ

um]

0044

43JK

973679

475

ProteinSE

T-lik

e[Brachypodium

dista

chyon]

7119890minus33

44JK

973680

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

45JK

973681

389

Predictedprotein[H

ordeum

vulga

re]

4119890minus27

46JK

973682

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

47JK973683

210

Putativ

ecalcium

exchanger[Triticum

dicoccoides]

4119890minus27

48JK

973684

478

Thioredo

xin[M

edica

gotru

ncatula]

2119890minus75

49JK

973685

210

Nosignific

antm

atch

50JK

973686

553

Root

nodu

leextension[Pisu

msativa]

5119890minus04

51JK

973687

600

Sign

alrecogn

ition

particleprotein[O

ryza

sativa]

5119890minus08

52JK

973688

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

53JK

973689

395

MLO

-like

protein[M

edica

gotru

ncatula]

2119890minus24

54JK973690

250

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

55JK973691

215

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

56JK

973692

411

Unk

nown[G

lycinem

ax]

11

57JK973693

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

58JK973694

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

59JK973695

420

Uncharacterized

protein[Zea

mays]

9119890minus83

60JK

973696

350

Uncharacterized

protein[Zea

mays]

5119890minus51

61JK

973697

244

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

5119890minus27

62JK

973698

415

Os01g0678900[O

ryza

sativa]

8119890minus11

63JK

973699

282

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]3119890minus51

64JK

973700

558

60Srib

osom

alprotein[Vitisv

inifera]

8119890minus65

65JK

973701

490

Nosignific

antm

atch

66JK

973702

556

Root

nodu

leextension[Pisu

msativ

um]

3119890minus07

67JK

973703

313

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]2119890minus65

68JK

973704

311

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]1119890minus64

69JK

973705

320

Hypothetic

alproteinMTR

4g076190

[Medica

gotru

ncatula]

2119890minus13

70JK

973706

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

um]

9119890minus119

71JK

973707

460

Transm

embranee

mp24do

main-containing

protein10-like

[Brachypodium

dista

chyon]

2119890minus45

72JK

973708

482

60Srib

osom

alprotein[Zea

mays]

3119890minus52

73JK

973709

395

MLO

5-lik

eprotein

[Medica

gotru

ncatula]

2119890minus24

74JK

973710

480

Type

2metallothionein

[Prosopisjulifl

ora]

4119890minus28

75JK

973711

556

Root

nodu

leextension[Pisu

msativa]

3119890minus07

76JK

973712

174

Hypothetic

alprotein[O

ryza

sativa]

086

77JK

973713

278

Nosignific

antm

atch

78JK

973714

439

Nosignific

antm

atch

79JK

973715

439

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

2119890minus31

80JK

973716

551

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

9119890minus39

81JK

973717

347

Nosignific

antm

atch

82JK

973718

661

Gam

ma-glutam

ylhydrolase[Vitis

vinifer

a]1119890minus54

83JK

973719

499

Type

2metallothionein

[Prosopisjuliflora]

2119890minus30


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

84JK

973720

463

Nod

ulin

mtn21eam

a-lik

etranspo

rter


]2119890minus44

85JK

973721

200

RNArecogn

ition

motif-containing


]4119890minus10

86JK

973722

280

Beta-glucosid

ase4

4-lik

e[Glycinem

ax]

3119890minus52

87JK

973723

424

V-type

proton

ATpase

21kD

Aproteolip

idsubu

nit[Medica

gotru

ncatula]

2119890minus29

88JK

973724

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

89JK

973725

540

Polyrib

onucleotiden

ucleotidyltransfe

rase

[Vitisv

inifera]

3119890minus07

90JK

973726

649

Methion

inea

minop

eptid

ase2

B-lik

e[Brachypodium

dista

chyon]

3119890minus40

91JK

973727

585

Hypothetic


69

92JK

973728

487

Unk

nown[Lotus

japonica]

1

93JK

973729

558

60Srib

osom


8119890minus65

94JK

973730

314

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]1119890minus55

95JK

973731

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

96JK

973732

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

97JK

973733

239

NOT2

NOT3

NOT5

family

protein[O

ryza

sativa]

3119890minus62

98JK

973734

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

99JK

973735

559

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

100

JK973736

793

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

101

JK973737

649

Methion

inea

minop

eptid

ase2

B[Brachypodium

dista

chyon]

1119890minus24

102

JK973738

210

Gam


uscommun

is]7119890minus10

103

JK973739

553

Unk

nownproteinprod

uct[Glycinem

ax]

1119890minus04

104

JK973740

600

Glycine


ax]

4119890minus102

105

JK973741

572

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04




4 Conclusion


Abbreviations





References

























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table3Hom

olog

ysearch

oftheE

STsg

enerated

from

pigeon

pear

ootalong

with

their119864

valueu

singBL

AST

xprogramme

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

1JK

973637

560

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

2JK

973638

231

Mito

chon

drialinn

ermem

branep

rotein

OXA1-like[

Vitis

vinifer

a]5119890minus41

3JK

973639

630

Uncharacterized


]9119890minus159

4JK

973640

621

Hypothetic

alprotein[Sorghum

bicolor]

9119890minus139

5JK

973641

570

GTP

-binding

proteinhfl

x[M

edica

gotru

ncatula]

6119890minus13

6JK

973642

412

Unk

nown[G

lycinem

ax]

11

7JK

973643

264


mem

branep

rotein

3-lik

e[Vitis

vinifer

a]2119890minus55

8JK

973644

438

Histon

e2[Populus

trichocarpa]

5119890minus63

9JK

973645

200

RNArecogn

ition

motif-containing


]4119890minus10

10JK973646

870

Beta-glucosid

ase[

Arabidopsis

thaliana

]00

11JK

973647

555

Epsin

N-te

rminalho

molog

y(ENTH

)dom

ain-containing

protein[M

edica

gotru

ncatula]

4119890minus36

12JK

973648

210

Gam


uscommun

is]7119890minus10

13JK

973649

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

14JK

973650

474

Glycine


chon

driallike)[G

lycinem

ax]

4119890minus102

15JK

973651

432

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

16JK

973652

227

TBP-associated

factor


]0051

17JK

973653

290

TBP-associated

factor


]8119890minus48

18JK

973654

303

Transcrip

tionfactor

[Medica

gotru

ncatula]

6119890minus29

19JK

973655

432

Uncharacterized

protein[G

lycinem

ax]

8119890minus11

20JK

973656

237

Nuclear

cap-bind

ingproteinsubu

nit[Arabidopsis

thaliana

]8119890minus48

21JK

973657

501

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04

22JK

973658

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

23JK

973659

560

Two-compo

nent

respon

seregulator

ARR

9[Vitisv

inifera]

1119890minus45

24JK

973660

210

Ribo

nucle

oside-diph

osph

ater

eductase

smallchain

like[Glycinem

ax]

2119890minus34

25JK

973661

748

Hypothetic

alprotein[O

ryza

sativa]

26JK

973662

742

Cyclin-depend

entp

rotein

kinase

complex

compo

nent

[Aspergillu

skaw

achii]

012

27JK

973663

406

Elon

gatio

nfactor

2[N

icotia

natobacum]

5119890minus39

28JK

973664

841

Expressedprotein[O

ryza

sativa]

73

29JK

973665

385

Hypothetic

alprotein[Vitisv

inifera]

21

30JK

973666

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

31JK

973667

510

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

6119890minus77

32JK

973668

630

Nosignific

antm

atch

33JK

973669

804

Gag-pro[Pisu

msativ

um]

9119890minus18

34JK

973670

320


[Medica

gotru

ncatula]

2119890minus13

35JK

973671

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

m]

6119890minus114

36JK

973672

371

ABC

transporterF

family

mem

ber1-like

[Brachypodium

dista

chyon]

4119890minus60

37JK

973673

513

Predictedprotein[H

ordeum

vulga

re]

2119890minus89

38JK973674

465

Phosph


maestivum]

2119890minus97

39JK973675

465

Phosph


maestivum]

2119890minus97

40JK

973676

457

Unk

nown[Pice

asitchensis]

097

41JK

973677

785

GTP

-binding

signalrecognitio

nparticleSR

P54[M

edica

gotru

ncatula]

7119890minus51


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

42JK

973678

241

Sign

alrecogn

ition

particle54

kDAsubu

nitp

recursor

[Pisu

msativ

um]

0044

43JK

973679

475

ProteinSE

T-lik

e[Brachypodium

dista

chyon]

7119890minus33

44JK

973680

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

45JK

973681

389

Predictedprotein[H

ordeum

vulga

re]

4119890minus27

46JK

973682

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

47JK973683

210

Putativ

ecalcium

exchanger[Triticum

dicoccoides]

4119890minus27

48JK

973684

478

Thioredo

xin[M

edica

gotru

ncatula]

2119890minus75

49JK

973685

210

Nosignific

antm

atch

50JK

973686

553

Root

nodu

leextension[Pisu

msativa]

5119890minus04

51JK

973687

600

Sign

alrecogn

ition

particleprotein[O

ryza

sativa]

5119890minus08

52JK

973688

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

53JK

973689

395

MLO

-like

protein[M

edica

gotru

ncatula]

2119890minus24

54JK973690

250

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

55JK973691

215

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

56JK

973692

411

Unk

nown[G

lycinem

ax]

11

57JK973693

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

58JK973694

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

59JK973695

420

Uncharacterized

protein[Zea

mays]

9119890minus83

60JK

973696

350

Uncharacterized

protein[Zea

mays]

5119890minus51

61JK

973697

244

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

5119890minus27

62JK

973698

415

Os01g0678900[O

ryza

sativa]

8119890minus11

63JK

973699

282

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]3119890minus51

64JK

973700

558

60Srib

osom

alprotein[Vitisv

inifera]

8119890minus65

65JK

973701

490

Nosignific

antm

atch

66JK

973702

556

Root

nodu

leextension[Pisu

msativ

um]

3119890minus07

67JK

973703

313

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]2119890minus65

68JK

973704

311

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]1119890minus64

69JK

973705

320

Hypothetic

alproteinMTR

4g076190

[Medica

gotru

ncatula]

2119890minus13

70JK

973706

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

um]

9119890minus119

71JK

973707

460

Transm

embranee

mp24do

main-containing

protein10-like

[Brachypodium

dista

chyon]

2119890minus45

72JK

973708

482

60Srib

osom

alprotein[Zea

mays]

3119890minus52

73JK

973709

395

MLO

5-lik

eprotein

[Medica

gotru

ncatula]

2119890minus24

74JK

973710

480

Type

2metallothionein

[Prosopisjulifl

ora]

4119890minus28

75JK

973711

556

Root

nodu

leextension[Pisu

msativa]

3119890minus07

76JK

973712

174

Hypothetic

alprotein[O

ryza

sativa]

086

77JK

973713

278

Nosignific

antm

atch

78JK

973714

439

Nosignific

antm

atch

79JK

973715

439

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

2119890minus31

80JK

973716

551

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

9119890minus39

81JK

973717

347

Nosignific

antm

atch

82JK

973718

661

Gam

ma-glutam

ylhydrolase[Vitis

vinifer

a]1119890minus54

83JK

973719

499

Type

2metallothionein

[Prosopisjuliflora]

2119890minus30


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

84JK

973720

463

Nod

ulin

mtn21eam

a-lik

etranspo

rter


]2119890minus44

85JK

973721

200

RNArecogn

ition

motif-containing


]4119890minus10

86JK

973722

280

Beta-glucosid

ase4

4-lik

e[Glycinem

ax]

3119890minus52

87JK

973723

424

V-type

proton

ATpase

21kD

Aproteolip

idsubu

nit[Medica

gotru

ncatula]

2119890minus29

88JK

973724

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

89JK

973725

540

Polyrib

onucleotiden

ucleotidyltransfe

rase

[Vitisv

inifera]

3119890minus07

90JK

973726

649

Methion

inea

minop

eptid

ase2

B-lik

e[Brachypodium

dista

chyon]

3119890minus40

91JK

973727

585

Hypothetic


69

92JK

973728

487

Unk

nown[Lotus

japonica]

1

93JK

973729

558

60Srib

osom


8119890minus65

94JK

973730

314

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]1119890minus55

95JK

973731

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

96JK

973732

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

97JK

973733

239

NOT2

NOT3

NOT5

family

protein[O

ryza

sativa]

3119890minus62

98JK

973734

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

99JK

973735

559

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

100

JK973736

793

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

101

JK973737

649

Methion

inea

minop

eptid

ase2

B[Brachypodium

dista

chyon]

1119890minus24

102

JK973738

210

Gam


uscommun

is]7119890minus10

103

JK973739

553

Unk

nownproteinprod

uct[Glycinem

ax]

1119890minus04

104

JK973740

600

Glycine


ax]

4119890minus102

105

JK973741

572

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04




4 Conclusion


Abbreviations





References

























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

42JK

973678

241

Sign

alrecogn

ition

particle54

kDAsubu

nitp

recursor

[Pisu

msativ

um]

0044

43JK

973679

475

ProteinSE

T-lik

e[Brachypodium

dista

chyon]

7119890minus33

44JK

973680

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

45JK

973681

389

Predictedprotein[H

ordeum

vulga

re]

4119890minus27

46JK

973682

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

47JK973683

210

Putativ

ecalcium

exchanger[Triticum

dicoccoides]

4119890minus27

48JK

973684

478

Thioredo

xin[M

edica

gotru

ncatula]

2119890minus75

49JK

973685

210

Nosignific

antm

atch

50JK

973686

553

Root

nodu

leextension[Pisu

msativa]

5119890minus04

51JK

973687

600

Sign

alrecogn

ition

particleprotein[O

ryza

sativa]

5119890minus08

52JK

973688

544

ADP-rib


8d[N

icotia

natobacum]

2119890minus105

53JK

973689

395

MLO

-like

protein[M

edica

gotru

ncatula]

2119890minus24

54JK973690

250

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

55JK973691

215

Phosph


rotein

[Ricinu

scom

mun

is]3119890minus09

56JK

973692

411

Unk

nown[G

lycinem

ax]

11

57JK973693

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

58JK973694

246

Phosph


rotein

[Ricinu

scom

mun

is]5119890minus09

59JK973695

420

Uncharacterized

protein[Zea

mays]

9119890minus83

60JK

973696

350

Uncharacterized

protein[Zea

mays]

5119890minus51

61JK

973697

244

Jmjcdo

main-containing

protein4-lik

e[Brachypodium

dista

chyon]

5119890minus27

62JK

973698

415

Os01g0678900[O

ryza

sativa]

8119890minus11

63JK

973699

282

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]3119890minus51

64JK

973700

558

60Srib

osom

alprotein[Vitisv

inifera]

8119890minus65

65JK

973701

490

Nosignific

antm

atch

66JK

973702

556

Root

nodu

leextension[Pisu

msativ

um]

3119890minus07

67JK

973703

313

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]2119890minus65

68JK

973704

311

Coiled-coildo

main-containing

protein[Ricinu

scom

mun

is]1119890minus64

69JK

973705

320

Hypothetic

alproteinMTR

4g076190

[Medica

gotru

ncatula]

2119890minus13

70JK

973706

509

Putativ

eS-adeno

sylm

ethion

ines

ynthetase[Ca

psicu

mannu

um]

9119890minus119

71JK

973707

460

Transm

embranee

mp24do

main-containing

protein10-like

[Brachypodium

dista

chyon]

2119890minus45

72JK

973708

482

60Srib

osom

alprotein[Zea

mays]

3119890minus52

73JK

973709

395

MLO

5-lik

eprotein

[Medica

gotru

ncatula]

2119890minus24

74JK

973710

480

Type

2metallothionein

[Prosopisjulifl

ora]

4119890minus28

75JK

973711

556

Root

nodu

leextension[Pisu

msativa]

3119890minus07

76JK

973712

174

Hypothetic

alprotein[O

ryza

sativa]

086

77JK

973713

278

Nosignific

antm

atch

78JK

973714

439

Nosignific

antm

atch

79JK

973715

439

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

2119890minus31

80JK

973716

551

Serin

ecarbo

xypeptidase-lik

e19-lik

e[Glycinem

ax]

9119890minus39

81JK

973717

347

Nosignific

antm

atch

82JK

973718

661

Gam

ma-glutam

ylhydrolase[Vitis

vinifer

a]1119890minus54

83JK

973719

499

Type

2metallothionein

[Prosopisjuliflora]

2119890minus30


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

84JK

973720

463

Nod

ulin

mtn21eam

a-lik

etranspo

rter


]2119890minus44

85JK

973721

200

RNArecogn

ition

motif-containing


]4119890minus10

86JK

973722

280

Beta-glucosid

ase4

4-lik

e[Glycinem

ax]

3119890minus52

87JK

973723

424

V-type

proton

ATpase

21kD

Aproteolip

idsubu

nit[Medica

gotru

ncatula]

2119890minus29

88JK

973724

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

89JK

973725

540

Polyrib

onucleotiden

ucleotidyltransfe

rase

[Vitisv

inifera]

3119890minus07

90JK

973726

649

Methion

inea

minop

eptid

ase2

B-lik

e[Brachypodium

dista

chyon]

3119890minus40

91JK

973727

585

Hypothetic


69

92JK

973728

487

Unk

nown[Lotus

japonica]

1

93JK

973729

558

60Srib

osom


8119890minus65

94JK

973730

314

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]1119890minus55

95JK

973731

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

96JK

973732

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

97JK

973733

239

NOT2

NOT3

NOT5

family

protein[O

ryza

sativa]

3119890minus62

98JK

973734

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

99JK

973735

559

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

100

JK973736

793

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

101

JK973737

649

Methion

inea

minop

eptid

ase2

B[Brachypodium

dista

chyon]

1119890minus24

102

JK973738

210

Gam


uscommun

is]7119890minus10

103

JK973739

553

Unk

nownproteinprod

uct[Glycinem

ax]

1119890minus04

104

JK973740

600

Glycine


ax]

4119890minus102

105

JK973741

572

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04




4 Conclusion


Abbreviations





References

























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table3Con

tinued

Slnum

ber

Gen

bank

accessionnu

mber

Leng

th(in

bp)

Hom

ologou

sprotein

119864-value

84JK

973720

463

Nod

ulin

mtn21eam

a-lik

etranspo

rter


]2119890minus44

85JK

973721

200

RNArecogn

ition

motif-containing


]4119890minus10

86JK

973722

280

Beta-glucosid

ase4

4-lik

e[Glycinem

ax]

3119890minus52

87JK

973723

424

V-type

proton

ATpase

21kD

Aproteolip

idsubu

nit[Medica

gotru

ncatula]

2119890minus29

88JK

973724

210

AP2

ERF

andB3

domain-containing

transcrip

tionfactor


]4119890minus40

89JK

973725

540

Polyrib

onucleotiden

ucleotidyltransfe

rase

[Vitisv

inifera]

3119890minus07

90JK

973726

649

Methion

inea

minop

eptid

ase2

B-lik

e[Brachypodium

dista

chyon]

3119890minus40

91JK

973727

585

Hypothetic


69

92JK

973728

487

Unk

nown[Lotus

japonica]

1

93JK

973729

558

60Srib

osom


8119890minus65

94JK

973730

314

D-3-Pho

spho

glycerated

ehydrogenase

[Ricinu

scom

mun

is]1119890minus55

95JK

973731

350

EIN3-bind

ingF-bo


dista

chyon]

8119890minus65

96JK

973732

499

DNA-

bind

ingproteinRA

V1[Zeamays]

4119890minus25

97JK

973733

239

NOT2

NOT3

NOT5

family

protein[O

ryza

sativa]

3119890minus62

98JK

973734

272

40Srib

osom

alproteinS24-2-lik

e[Brachypodium

dista

chyon]

2119890minus57

99JK

973735

559

Sign

alrecogn

ition

particlesubu

nit[Arabidopsis

thaliana

]9119890minus80

100

JK973736

793

Vesic

le-associatedmem

branep

rotein

727-lik

e[Glycinem

ax]

1119890minus111

101

JK973737

649

Methion

inea

minop

eptid

ase2

B[Brachypodium

dista

chyon]

1119890minus24

102

JK973738

210

Gam


uscommun

is]7119890minus10

103

JK973739

553

Unk

nownproteinprod

uct[Glycinem

ax]

1119890minus04

104

JK973740

600

Glycine


ax]

4119890minus102

105

JK973741

572

Glucanendo


eprotein

3-lik

e[Glycinem

ax]

2119890minus04




4 Conclusion


Abbreviations





References

























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




4 Conclusion


Abbreviations





References

























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Research Article Identification and Validation of ...downloads.hindawi.com/archive/2014/651912.pdfInternationalJournalofPlantGenomics T :SummaryofESTslibrary. Total clone sequenced