Acclimatization of Bambusa

8/14/2019 Acclimatization of Bambusa

http://slidepdf.com/reader/full/acclimatization-of-bambusa 1/4

© SocietyofApplied Sciences

Acclimatization and Macroproliferation of Micropropagated Plants of Roxb. Bambusa tulda

Yogeshwar Mishra*, Pradeep Patel and S.A.AnsariGenetics and Plant Propagation Division

Tropical Forest Research Institute, P.O.-R.F.R.C., Mandla Road, Jabalpur (M.P) 482021, India

INTRODUCTIONMicropropagation has been widely utilized for rapid and mass multiplication of many species including bamboos.However, its sample application is restricted often by the high rate of percentage of plant loss when transferred to

condition.Thisis due to regenerates get adjusted withmanyabnormalities in environment like high levelof irradiance, lowhumidity andwater is limiting dueto lowhydraulicconductivityof roots androot-stemconnections[1]. Acclimatization of regenerates will overcome these problems with gradual lowering in air humidity [2]. Thedevelopment of successful hardening technique is prerequisite for micropropagation method. These days manycomputer based acclimatization units are available but at a high price [3]. Hardening accounts for about 60% of thetotal production cost [4]. Therefore, an efficient and cost-effective acclimatization technique is necessary for raised plantlets. So, we attempted to develop a new efficient, cost-effective and one step of hardeningtechnique for raised plantlets of Roxb. It is a very versatile and rapidly growing bamboo of South East Asia but suffer from enormous reduction in their stocks due to over exploitation by ever-increasing

population and colossal use in pulp and paper industries. is a native to north and north-east India, Bangladesh,Myanmar and Thailand [5]. It has been reported as a reluctant-to-root species [6, 7]. Its culms are used for construction, scaffolding, furniture, handicrafts and paper pulp while young shoots are pickled [8]. Being rich in

phytosterols, shoots are excellent source for production of sterol drugs [9]. Various tissue culture protocols

have been evolved for large scale production of the plantlets of a using seedling and field grown explants [10-12]. However, hardening of plantlets remains a crucial step for success of tissue culture protocols of thisspecies. No attempt has been made describing detailed hardening procedure of this species. In the present paper, theauthors describe a method for acclimatization and successful hardening of the plantlets of raisedthrough micropropagation. Following the pioneering work done by Banik [13] andAdarsh Kumar . [14], we haveoptimized the macroproliferation technique of hardened and acclimatized plantlets of for further stock buildup.

exvitro ex vitro

in vitroin vitro

in vitro Bambusa tulda

B. tulda

B. tulda

B.tuld in vitro

Bambusa tuldaet al

B. tulda

ASIAN J. EXP. BIOL. SCI. VOL 2(3) 2011: 498-501

SHORT COMMUNICATION

ABSTRACT

KEY WORDS:

In the present paper, a procedure for acclimatization and macroproliferation of in vitro plantlets raised from field grownclump of Bambusa tulda is described. The acclimatization procedure included gradual hardening from culture room tonatural environment, taking 30 – 45 days. Rooted plantlets of about 2-3 cm height were subsequently transferred to different

potting media for acclimatization. Plantlets grown in perlite showed 91% survival followed by soilrite with 66% of survival.Theacclimatized plantletsweremacroproliferated in a cycle of sixmonths andprovided with a uniform dose of 100ppmurea.Theproliferation ratewasstabilized to be 3 plantletspermacroproliferation cycle in B.tulda.

Hardening, MS inorganic salts, plantlets production, substratum

ASIAN J. EXP. BIOL. SCI .VOL 2(3) 2011498



MATERIALSAND METHODSHardeningexperiment

In vitro B. tulda

in vitro

ex vitro

regeneration of plantlets was achieved by the method described earlier [12]. The plantlets of 2-4 cmlength were taken out from the culture flasks and washed in running tap water thoroughly so as to remove adheredmedium from the surface of plantlets followed by washings with 0.1% (w/v) Bavistin® for 2 min and tap water. Thewashed plantlets were transferred to root trainers comprising 25 cells each of 150cc filled with four different potting

media viz., soilrite, perlite, vermiculite and compost for hardening (Fig 1a). The potting media were autoclaved at121 C of1.05kg cm for 1 hrbefore being used for hardening.

The planted root trainers shifted to plastic tray filled with half strength of iron free MS salts covered with perforatedtransparent polythene sheet and maintained at 24 ± 1 C under normal laboratory light condition (40 µ mol /m /sec.)

provided with fluorescent tube and 12/12 h (light/dark) photoperiod for hardening (Fig 1b). The observationswere recorded forsurvivalpercentage andheight of theplants after 8 weeks of transfer.After an acclimatizationperiodof 8 weeks when newleaveshademerged, theplantletswere transferred to shadehouse andmaintained for2 weeks for

hardening (Fig 1c). Finally, the hardened and acclimatized plantlets were transplanted in polythene bagscontaining soil: sand: farmyard manure (1:1:1) in theshadehouse condition for1½ to 2 monthsreceiving irrigation on

0 -1

0 2

Fig.1:Hardeningandmacroproliferationof raised Roxb.:inv itro B.tulda (a) transfer of plantletsin root trainer filled withdifferenthardeningsubstratum;(b) roottrainershifted totray filledwith½ MS solution andcovered with perforated polythenesheet; (c) hardened

plantlets ready for transfer to mist chamber; (d) macroproliferable plantlets after six month of transfer; (e-g) separation and planting of

individual tillers in polythene bags (h) macroproliferated propagules maintained in shadehouse; (i) six month old propagule ready for macroproliferation.

in vitro

Acclimatization ……………………….....................................................................................……. Roxb............Yogeshwar Mishra Bambusa tulda et al.

ASIAN J. EXP. BIOL. SCI .VOL 2(3) 2011 499



alternate daysand spray of½ MSsolutiononce in15 days.In the hardening experiment, a total of25 plants (in a root trainer) were taken for eachpotting medium and regarded asone replication. A total of three replications were used in this experiment. representing means of threereplications were analysed with the SX statistical package according to a completely randomize design (CRD) usingone way analysis of variance. Least Significant Differences (LSD) values were calculated at = 0.05 for comparingmeans of the treatments.

Thehardened andacclimatizedplantletswere planted in polybags of 24x18 cm size in themonth of October 2006. Sixmonths old hardened plantlets of having tillers, rhizomes and roots were carefully removed from the polybags(Fig 1d). Each proliferated tiller along with some rhizome and roots was separated to act as a propagule (Fig 1e, f) and

planted in fresh polythene bags filled with soil: sand: farmyard manure (1:1:1) (Fig 1g) in the month of March andinitially supplied with 100 ppm urea and maintained in shadehouse (Fig 1h). The data on average diameter of rhizomes, average root length, average root number, average tiller diameter, average number of nodes, average tiller lengthand average numberof tillers were recorded after sixmonths of transfer (Fig 1i).

representing means of three replications were analysed with the SX statistical package according to a completelyrandomize design (CRD) using one-way analysis of variance. Least Significant Differences (LSD) values werecalculated at = 0.05 forcomparingmeansof thetreatments.

The data recorded on the survival percentage and height of the plants during hardening is presented in Table 1. In thehardening experiment, perlite resulted in maximum survival (91%) of plantlets of , which was statisticallyhigher than all other substrates. Inaccordance withthisfinding, Ndiaye . [15]andLin and [16] were alsofound perlite as one of the most suitable potting media for hardening of bamboo species. Macroproliferation is asimple and effective procedure of vegetative propagation recommended when bamboo plants are available. Thistechnique was successfullyusedbyBanik [17],AdarshKumar . [18]andAdarsh Kumar . [19]and attemptedatTropicalForestResearch Institute, Jabalpur. In this method, thetillers (individual thin culms along with rhizome) of a proliferated seedling were separated (by rhizome division) and planted as individual propagules. New tillers arosefrom these propagules in a few months. When they reached a 3–4 tiller stage, they were again subjected to tiller separation and planting. The regenerated plantlets subjected to macroproliferation produced 3 and 2.44number of tillers in inMarch –August and May-October, respectively (Table 2). The data revealed that there is

significant variation in number of tillers in this species in two seasons. The relatively low number in May-October maybe possibly due to theonset of mansonandcomparatively lowtemperature.Whereas there wasno dormant phase between March- August and the growth of propagule was steady and continuous. Accordingly, each propagule became suitable to give a maximum of 3 numbers of propagules in the month of March-August. Similarly, other parameters like, diameter of rhizome, root length, root number, tiller diameter and number of nodes are alsosignificantly higher in March-August season in as compared to May-October. In earlier study Banik [13]obtained 3 numbers of tillers after 9 month in macroproliferated seedling of . However, obtaining 3numbers of tillers in a shorter span of 6 months is an added advantage of this study, which may be attributed to the useof clonalmaterialsfor macroproliferation. Thevegetative propagationof through macroproliferationhas beenfound as one of the most dependable techniques. The multiplied tillers, thus produced, remain smaller in size due tocontinuous rhizome separation. This is additional advantageas such propagules are easy to handleand transport. This

procedure can be successfully applied to self incompatible bamboo particular that has inherent difficulty ingeneratingplants through vegetativemethods in general.

p

B.tulda

p

B.tuldaetal

et al et al

in vitro B.tulda

B.tulda Bambusa tulda

B.tulda

B.tulda

Macroproliferation experiment

RESULTSAND DISCUSSION

Table 1: Performance of raised plants of under different hardening substratum. Observation recordedafter 8 weeks

in vitro B.tulda

The data

A total of 30 tillers were taken and regarded as one replication. A total of three replications were used. The data

Chang


ASIAN J. EXP. BIOL. SCI .VOL 2(3) 2011500

ParametersPott ing mediaSurv ival % H eight (cm ) N ew leaf

formation (%)D a y s r eq u ir ed f or

leaf formationSoilrite 66 .6 (56 .6 ) 16 .2 76 12

Perlite 91 .0 (80 .0 ) 22 .33 89 9V erm iculite 58 .0 (50 .0 ) 21 .33 65 14C o m post 20 .0 (26 .56) 19 .67 15 16LSD (0 .05) 22 .67 6 .71 19 .23



CONCLUSION

REFERENCES

Theonestep hardeningtechnique describedherein provesto be cost effectiveandefficient alternative toconventionalhardening technique for acclimatization of raisedplantletsof . Productionof plantsin shorter period(6months) through macroproliferation procedure is an addedadvantageto produce clonal plantingmaterialsof raised through micropropagation.

in vitro B.tulda B.tulda

[1]. Fila, G.,Ghashghaie,J. andCornic, G. (1998). Photosynthesis, leaf conductance and water relation of in vitro cultured grapevinerootstock inrelationtoacclimatization, 102:411-418.

[2]. Lavanya, M., Venkateshwarlu, B. and Devi,B.P. (2009).Acclimatization of neem microshoots adaptable to semi sterile conditions,., 8:218-222.

[3]. Pospisilova, J., Ticha, I., Kadleaeek, P., Haisel, D. and Plazkova, S. (1999). Acclimatization of micropropagated plants toconditions. ., 42:481-497.

[4]. Hazarika, B.N.(2003).Acclimatization of raisedplants, 85: 1704-1712.[5]. Gamble, J. S. (1896).Bambusae of British India.Ann.Bot. Garden Calcutta,3:32-33.[6]. McClure, F.A.and Kennard,W.C. (1955). Propagation studies (withbamboo).Report of Federal Experimental Station Puerto Rico.[7]. Rao, I.V.R., Rao, U.I. and Roohi, F.N. (1992). (Ed. Baker, F.W.G.)

Rapid Propagation of Fast-growing Woody Species,CASAFAReport Series No.3: 41-56.[8]. Seethalakshmi,K.K.and Kumar, M.S.M.(1998). BambooInformationCentre-India, KFRI,Peechi.[9]. Srivastava, R.C.(1990). Bamboo, new raw materialforphytosterols. , 59: 1333-1334.[10].Saxena,S . (1990). propagationof thebamboo Roxb.) throughshootproliferation, ., 9 :431–434.[11]. Pal, A. and Das, M. (2005). Clonal propagation and production of genetically uniform regenerants from axillary meristems of adult

bamboo. , 14: 185-188.[12]. Mishra, Y., Patel, P., Yadav, S., Shirin, F. and Ansari, S.A. (2008) A micropropagation system for cloning of Roxb.

115:315-318.[13]. Banik, R.L. (1987). Techniques of bamboo propagation with special reference to pre-rooted and pre-rhizomed branch cuttings and tissue

culture (Ed. Rao, A. N.), IDRC, Canada, pp. 160–169.[14].Adarsh, Kumar., Gupta, B.B. and Negi, D.S. (1988). Vegetative propagation of through macroproliferation.

114:564–568.[15]. Ndiaye, A., Diallo, M.S., Niang, D. and Gassama-dia, Y.K. (2006). regeneration of adult trees of

.,5: 1245-1248.[16]. Lin, C.S. and Chang, W.C. (1998) Micropropagation of through nodal explants of field-grown culms and flowering of

regeneratedplantlets, .,17:617-620.[17]. Banik, R. L. (1995). INBARTechnical Report No. 6, INBAR, FORTIP and Bangladesh

, pp. 1–66.[18].AdarshKumar, Gupta,B.B. andNegi, D.S.(1991). Vegetative propagationof throughmacroproliferation II.

.,117: 621–623.[19].Adarsh Kumar, Pal, M. and Kumar, S. (1992).Mass production of field planting stock of vegetatively throughmacrorpoliferation. 118:638–645.

Physiol.Plant., Ind. J.

Biotechnol ex vitro

Biol. Plant in vitro Curr. Sci.,

Bamboo propagation through conventional and in vitro techniques

Bamboo of India.Curr. Sci.

In vitro (Bambusa tulda Plant CellRep

J. Plant Biochem.and Biotech. Bambusa tulda Sci.

Horti.,

Recent Researchon Bamboos Dendrocalamus strictus Ind.

For., In vitro Bambusa vulgaris, African J.

Biotech Bambusa edulis

Plant CellRep A manual for vegetativepropagation of Bamboos,

ForestResearch Institute Dendrocalamus strictus Ind.

For Dendrocalamus hamiltonii Ind. For.,

Table 2: Propagule production and growth data of in vitro raised plantlets of B.tulda

Number of propaguleParametersMarch-Aug May-Oct LSD (0.05)

Diameter rhizome (cm) 1.24 0.90 0.313Root length (cm) 34.33 23.33 10.82

Root number (cm) 28.22 20.44 3.91Tiller dia. (cm) 0.85 0.49 0.179

Number of nodes 10.66 7.22 3.13Tiller length (cm) 110.89 76.66 14.83 Number of tiller 3 2.44 0.48


ASIAN J. EXP. BIOL. SCI .VOL 2(3) 2011 501

Corresponding author: Yogeshwar Mishra , Genetics and Plant Propagation Division, Tropical Forest Research Institute,P.O. -R.F.R.C., Mandla Road, Jabalpur (M.P) 482021, India, e -mail: [email protected]

Documents

Acclimatization of Bambusa