Progress and perspective of maize molecular breeding in China

Jiansheng LiNational Maize Improvement Center

China Agricultural University

Outline of Presentation Maize production in China Introduction of national maize

improvement center Progress of molecular breeding with

marker technology. Perspective of molecular breeding

Chinese maize belt

A slope long narrow regions from northeast to southwest

Each dot equal 50 thousands hectares

15.00

20.00

25.00

30.00

35.00

40.00

1978

1980

1985

1990

1991

1992

1993

1994

1995

1996

1997

1998

1999

2000

2001

2002

2003

2004

2005

2006

2007

2008

2009

Year

p

l

a

n

t

a

r

e

aRice wheat maize

Planting areas of three major crop in past three decades

Maize becomes the number one crop since 2007

Total output of three major crops, 2009

rice

maize

wheat

34% of total

cereals in China

Data from national bureau of statistics, 2010

163.9 MT

151.1 MT 195.1 MT

Consumption of maize in China

0

50

100

150

200

250

2000 2008 2015

KG

Kg/person/year

79

200

126

Trend of grain yield per hectare

Genetic improvement makes a great contribution to increase production

National Maize Improvement CenterGeneral Information

The center was founded by Agricultural Ministry of China in 1998. The government invested about US $

1.2 millions, another 1millin last year. The center is a national research,

service, and training organization。

Staff Member

Nine full professors Five associated professors Nine technicians About 20 Ph. D. students About 30 Mast D. students

Maize Research Building

4th floor: Molecular Genetics Labs

3rd floor: Genetic Engineering Labs

2nd floor: Quality Analysis Labs

1st floor: Seed Labs

3600 M2

Research Areas

Germplasm Enhancement Applied gene engineering Applied molecular genetics Maize Genomics Maize Breeding

Germplasm Enhancement

To create maize novel germplasm for hybrid maize breeding, such as high-oil, high-starch

To enhance genetic basis of germplasm by introducing exotic germplasm

5个群体含油量选择比较

3

7

11

15

19

0 2 4 6 8 10 12 14 16 18

选择世代

含油量%

BHOAIHORYDSyn.D.OKYGHO

Fig. Development of High oil maize populations

PCA plot of four subpopulations in 527 lines

-0.3

-0.2

-0.1

0

0.1

0.2

0.3

-0.9 -0.7 -0.5 -0.3 -0.1 0.1 0.3

PC 2

(6.9

%)

PC1 (18.2%)

SS

NSS

TST

Mixed

527 maize lines (CIMMYT 235; China 238; GEM 54)GoldenGate Array-1536 SNPs

Applied Genetic Engineering

A high-efficiency system for maize transformation has been established.

The Bt gene, being resistant to maize borer, has been transferred into elite inbred lines.

Line with BT CK

CaMV35S intron BT Cry1Ac-M ORF NOSATG TGA

Transgenetic line with BT

Application molecular genetics

To map some interesting genes and QTL which control important traits with molecular markers. To develop molecular markers for

marker assistant selection in breeding.

Traits Publication

Agronomy Plant height Euphytica 155:117-124

Grain yield Molecular Breeding, 20：41-51

Disease resistance

Resistance to stalk rotResistance to head smut

TAG 117:1241–1251

Quality High oil TAG 120:665–678

Pro-VA Nature Genetics, 42: 322–327

Forage Quality

Crop Science.

Mapped QTL for important traits in center

Maize Genomics

Widely variation of gene numbers among inbreds

Study on the structure and function of centromere on maize chromosome

Hybrid maize breeding

The conventional breeding methods combined with marker technology are becoming major approaches.

The traits breeders focus on are grain yield, stress tolerances, and maturity.

Planting areas of hybrids released from NMIC

0 5000 10000 15000 20000 25000 30000

2007

2006

2005

2004

2003

2002

2001

2000

1999

1998

年份

累计推广面积（单位：万亩）

In past decay, hybrids released from NMIC were extended for 200 m hectares per year on average in China.

years

Accumulated planting areas of hybrids released from center

High-oil hybrid (oil content 8.2%)

Progress of molecular breeding in maize

A project of maize molecular breeding was funded by S&T ministry of China, 2006-2010.

Institutes: CAU, CAAS, Sichuan AU, Henan AU, Jilin CAAS, Shandong CAAS.

Purpose: to develop molecular markers associated with agronomy traits, and to use them to improve hybrids

LCYElycopene

δ-carotene

α-carotene

LCYB

crtRB3

zeinoxanthin

lutein

CYP97C

ABA

crtRB1

LCYB

β-cryptoxanthin

γ-carotene

β-carotene

LCYB

zeaxanthin

crtRB

GGPP

PSYPDSZ-ISOZDS/CRTISO

Harjes C et al, 2008, Science

Fu ZY (付志远）TAG, 2010

Yan JB and Kandianis C et al, 2010, Nature Genetics

Zhou Yi (周毅）

I I D D D D D D

PSY1 5’InDel

LCYE

crtRB1

PSY1

crtRB1 5’TE crtRB1 D4 crtRB1 3’TE

PSY1 SNP7

341bp293bp

48bp

crtRB3 SNP2239

PCR based friendly markers are available for the genes

A single cross:DE3/By815

Kernel W/100 34gKernel No. 35Row No. 14.4Grain Yield 8.8T/hPro-VA 6.63 ug g-1

Functional markers for QPM

Phi057 5’CTCATCAGTGCCGTCGTCCAT3’5’CAGTCGCAAGAAACCGTTGCC3’

o2o2

O2O2

A major QTL-qLA1-1 for plant architecture on Chromosome 1

Donor: YU82 RP: CHANG7-2 CHANG 7-1 BC4F1

umc1292

umc22257.8

umc11661.5

umc22265.4

bnlg18036.7

dupssr24.3

qLA1-12.1

bnlg14843.2

bnlg4392.8

phi3390178.2

bnlg18664.3

umc21717.8

Chr.1 Chr.2 Chr.3 Chr.4 Chr.5

Chr.6 Chr.7 Chr.8 Chr.9 Chr.10

Ten QTL for nitrogen efficiency in maize genome

umc1422

bnlg1064umc2032umc1635umc1065

bnlg1329phi127bnlg1633mmc014

umc1536

bnlg1141umc1230bnlg1520umc1525umc1736bnlg1893umc1207phi42743qHSR1umc2184umc2077umc2214

QTL

Bin2.09

IDP171

b0351K12

Mo17 BAC

300kb ~50 genes

QTL analysis Fine-mapping

Major resistant QTL for Head Smut

umc1293

umc1319

umc2018

umc1863

bnlg1079

umc1336

rfg1umc2349

bnlg2336

umc1246

phi062

umc1053

umc2350

umc1115

umc1678

umc1280

Bin10.4

QTLAI857162SR

umc2349

umc1246

AI861111SR

umc2350

bnlg2336

AI8571626SR

1 3 5 7 9 11 13

phi062

QTL analysis

Fine-mapping

Major resistant QTL for Stalk Rot

High oil line 11.65%

Normal oil line 3.53%

High oil QTL mapping and breeding

B73/By804 RIL, F7

Phenotyping 223 F7:8 B73/By804 RIL

Two years (2005,2006),Beijing;

Materials

Marker NumberSSR 202STS 17 Candidate gene

CAPS 7 Candidate gene

SNP 2 Candidate gene

Total 228 Candidate gene

Figure Gas chromatographic profile of fatty acid composition in maize kernel of parents

B73 By804

chr 1 chr 2 chr 3 chr 4 chr 5umc2105umc2101

1.0

umc22566.2

bnlg114420.8

umc101221.6

phi03612.3

bnlg163824.6

umc16556.9

mmc01324.7

umc16832.4

phi0532.6

sad70047.4

dupssr234.1

gat510.1

umc24082.6

bnlg19714.2

bnlg16054.6

umc13990.5

phi04625.7

umc1491umc1253

0.7

umc10975.7

phi02419.2

umc203614.4

umc211512.8

umc144716.1

umc16925.7

umc23737.7

umc122110.5

umc202611.4

bnlg123723.5

f22a12.9

phi0857.8bnlg23058.7

umc214321.9

aca5.5

phi96100

umc126516.2

kt27.2

umc151832.3

umc142214.5

umc177616.6

umc203224.7

phi083bnlg108

3.2

u4

5.1

umc1635

0.1

umc1003phi092

2.9

umc1065

2.9

nc003

4.6

pap4

0.1

umc2372

1.7

C9_3

3.9

mmc0271

23.1

kass2

13.7

umc1551

11.9

bnlg1520

4.3

apat5

6.1

phi101049

21.7

umc1017

phi21398427.6

phi09624.0

phi0266.5

bnlg12173.0

bnlg17554.2

umc12999.0

m13.1

bnlg22919.2

bnlg11898.4

umc14669.7

dupssr280.1

bnlg21624.3

umc22868.8

bnlg10237.0

umc153217.6

umc164916.7

umc1177

bnlg1429

38.1

umc140320.2

fad837.0

mcat811.9

phi001umc1598

2.3

umc2217

5.2

bnlg1811

4.5

Y7AC

0.7

bnlg2086

7.9

bnlg1884

2.7

umc2112

10.7

umc2228

7.6

umc1626

15.5

umc2232

9.1

umc1988

5.9

umc1122

18.1

bnlg400

4.7

bnlg1556f6a

4.8

umc1955

23.4

umc2047

21.1

ols1

3.6

phi308707

8.1

umc1553

13.6

umc1605

18.5

umc2100

0.7

umc1725

5.9

phi064

11.5

oil16:0 18:0 18:1 18:2

phi118umc205310.3

umc133713.6

phi05014.4

umc13671.9

umc20160.7

bnlg16552.8

umc1336umc2180

0.2

phi062

3.5

MZETC34

2.0

umc2163umc1506

10.0

bnlg1082

9.1

umc2122

1.6

umc1933

8.8

umc1196

7.1

umc1645

15.2

umc1139umc2042

1.8

umc10756.2

umc130423.0

phi1196.2

bnlg208216.6

umc13600.4

bnlg18633.7

phi1151.1

phi1001750.3

umc1735phi121

1.5

bnlg2046

2.3

umc1130

2.7

umc1562

0.8

umc1141

14.2

CA25

0.1

umc1960

1.9

umc1149

0.1

sad2702

4.9

umc1724

10.0

umc1268

7.6

umc1673

14.6

umc1663

8.0

phi080

1.5

umc1069phi015

0.2

bnlg1131

4.4

umc2208phi07510.2phi1260.7umc175314.1zct4260.3umc23142.4bnlg3910.6umc11330.4umc23130.4umc11950.8YISSR0.2umc15955.9Q85.5umc19798.8nc0107.1umc110511.9umc17958.3umc20069.2umc23191.8phi1290.2bnlg16170.2umc16140.8acda60015.5nc01313.0bnlg17327.6umc21628.4phi29985213.7umc205925.9

umc1241umc1695

5.7

mmc01718.3

phi03430.9

umc14090.9

umc14330.6

bnlg17920.1

phi09118.0

atf20.7

umc111212.4

umc233223.1

phi3281750.8

dupssr131.9

CA334.3

kass5001B0.3

umc11257.6

umc219712.1

phi0825.4

kb1phi028

1.5

umc1588umc2362

1.1

phi017

1.2

Q2

0.7

umc1170

1.4

dupssr6

0.7

bnlg244

13.7

bnlg1401

0.7

umc1037

5.6

umc2213

5.3

umc2337

6.3

phi065

0.6

fae2

2.0

umc1258

0.9

umc1688

2.3

umc1743

0.4

umc1654

7.3

umc2119phi032umc1771

1.5

umc1519

0.2

umc1094

0.4

umc1078

0.4

umc1471

12.2

umc1794

6.0

bnlg1525

26.5

bnlg1904

37.2

chr 6 chr 7 chr 8 chr 9 chr 10

Results of Oil QTL mapping

18:2

16:018:018:1

oil18:2

16:018:018:1

oil18:2

16:018:018:1

oil

chr 9 chr 6 chr 1

LOD profiles of main QTL for fatty acid and oil concentrations

Pa9 (16:0) 45.5% st6-1 (18:0) 22.4% ol6 (18:1) 27.1%

li1-1 (18:2) 19.1% oil1 15.9%

Trait QTL action Numberh2(%)

range subtotal$ % of each trait16:0 major 1 42 42.0 53.9

minor 7 0.8-9.1 23.3 30.0epistatic 6 0.4-6.1 12.6 16.1total* 77.9

18:0 major 1 15.0 15 24.3minor 5 1.3-9.3 38.4 62.2epistatic 5 1.5-2.0 8.3 13.5total* 61.7

18:1 major 1 27.7 27.7 41.3minor 8 0.7-9.0 32.7 48.7epistatic 4 0.5-2.5 6.7 10.0total* 67.1

18:2 major 1 48.3 48.3 65.7minor 8 0.7-6.3 17.5 23.8epistatic 7 0.5-1.6 7.7 10.5total* 73.5

Oil major 1 15.7 15.7 26.0minor 8 1.5-8.4 39.6 65.4epistatic 2 1.3-3.9 5.2 8.6total* 60.5

Genetic Basis for Fatty Acid Compositions and Oil Content

Major and minor QTL as well as epistasiscontribute to fatty acid compositions and oilcontent in high-oil maize.

Three largest effect QTL were located onchromosome 1,6 and 9.

Conclusion of high oil QTL mapping

umc1177

bnlg1429

umc2217

bnlg2086

umc2232umc1122

umc1955

phi308707

phi064

0

38.1

74.7

87.8

133.4

157.4

190.3

223.1

273.3

Two Targeted Genomic Regions for MAS

Chromosome A (%) R2 (%）

Chr.1 0.48 15.7Chr.6 0.35 8.4

Chr.1umc2208

Y1SSR

Q8

bnlg1732

umc1979

umc1105

umc1614

phi299852

umc2059

0

30.1

41.5

115.9

50.3

69.3

89.8

138

163.9

Chr.6

By804 Zheng58 / Chang7-2X

F1 X

XBC1

BC2

BCn

X

Donor Recurrent Parents

MAS

Zheng58 / Chang7-2

Zheng58 / Chang7-2

Zheng58 / Chang7-2

Protocol for MAS-BC

ASK DGAT MVGNMIFWFFFSIVGQPMCVLLYEFO9B DGAT MVGNMIFWFF-SIVGQPMCVLLY

65.1

54.3

43.4

32.5

21.7

10.8

0.0

F value

DGAT1 - 2

DGAT1 - 2acyl-CoA:diacylglycerol acyltransferase

F469

Favorable AlleleTTC Insertion

Peizhong Zheng et. al, Nat Genet. 2008

1 15 30 33 45 60 63 75 81

Fig HO06 marker among high oil lines and normal lines

F469 code No F469 code

High oil 33（1-33）

Normal 18（64-81） 69份

Function marker for DGAT1-2

3.14

3.74

3.69

3.97

3.79

4.65

0

1

2

3

4

5

6

chang7-2 zheng58

oil co

nten

t (%

)

-/-

-/+

+/+

Variation of oil contents in BC6 F2

unc191

M53-1

M3

7K314

M5

M26-8

umc19

BC6_chang7-2_chr6 1 1 3 3 3 1 1

BC6_zheng58_chr6 1 1 3 3 3 1 1

3.82

4.65

0

1

2

3

4

5

6

-/- +/+

oil c

onte

nt (%

)

0

20

40

60

80

100

120

140

160

grain

weig

ht (g

)

oilcontent

grainweight

135 138

Oil content and grain weight in iso-genetic hybrids

Compared with normal Zhengdan958, the oil content increased 0.83%, grain weight had not changed

PerspectiveMolecular markers: From SSR to SNP,

High-through put SNP markers will be widely used.

Traits: From simple to complex,More complicated traits will be focused on for increasing grain yield.

Genome wide selection will be conducted in molecular breeding.

Challenge To train maize breeders with advanced

techniques in order to fill the gape between genetics and breeders.

To enhance the communication between public sections and seed industries in order to support the development of domestic seed companies.

AcknowledgeProfessor: DAI JR; XU ML; LAI JS;CHEN SJ; JIN WW

Dr. Jianbing Yan; Dr.Xiaohong Yang

863 High-Tech ProjectNature Science Fund of China