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ISO 9001 :2008 Orgonizotion
January-February 2016 Vegetables for Nutritional and Livelihood Security
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ISO 9001 :2008 Organization
Ho
e
Ja"uory~b.vory 2016
INDI.AN
o t·eD ture
January-February 2016 Published bimo thly, Vol. 61, No. 1
co Cover
From the Editor
: Vegetable wealth
Courtesy
Dr B. Singh, IIVR, Varanasi
EDITORIAL COMMITTEE Chairman N K Krishna Kumar Members S K Sharma
•
S K Malhotra
B N S Murthy
•
T Janakiram
Rameshwar Singh
•
S K Singh
Member-Secretory Sam Dutt
Editor
Art
Sam Dutt Norendra Bahadur
Design & Production: Punit Bhasin
Proiect Director (DKMA) : Rameshwar Singh Inchorge
(EEU)
: Aruna T Kumar
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Indian Council of Agricultural Research
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Microgreens are new generation smart food 3 TK. Kaley, Sudhir Singh, R.N. Prasad and B. Singh 5 Producing orgonic vegetables for bettsr health S.K. Singh, R.B. Yadava, S.N.S. Chaurasia R. N. Prasad, Raghwendra Singh, Paresh Chaukhande and B. Singh Quality seed of vegetables brings dividends 8 P.M. Singh, Manimurugan, T Chaubey, Rajesh Kumar, Vikas Singh and Nakul Gupta Genetically-modified vegetables for farmers' welfare 11 ( Maior Singh, Raiesh Kumar, H.C. Prasanna, S.K. Tiwari, R.S. Guiiar and S.G. Karkute Leafy vegetables for health and nutritional security 15 Pragya Ranjan, B.K. Singh, Y.S. Reddy and R.K. Dubey Genetic resources for future 18 B. Singh, M. Singh, H. Lal. N. Rai, DR. Bh loss) caused by brinjal fruit.- and shoot-borer (Leucinodes orbonalis) , Insecticides application to control FSB is oft n ineffecli e, incr'asing produ ti n costs and poses environm ntal hazards. a k of any natural urce of resi lance t.o FSB in all ulti 'aled and conventionally eros -compatible sp cies of Solanum have been major obstacles to developing F B-r-sistant cultivars. Cry proteins were permilted to develop Lepidopteran insect pest resistant crop plant.s. To d vel lp F B re istant b 'njal,
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ry7 c g ne wa transformed into an improved inbred lin 0 brinj I (IVBL-~)) via Agrobaclerium-mediat dune transfer. Nine single copy trans enie events wer d v loped. The quantilative EllS analyses estimat d ignifi a.nt Ie el of Cry7Ac protein (:2.4(;-4.33 nlY!ml I a e 'tra t) expr ssion in all the 9 independently tnmsfonnecl lines. Bio' sa s of transgenic (To) plants were conducted breI asing 7 da's-old pre-weighed FSB n onates on inta l sh Jots f ~O-days old seedlings and ;) rom-thick as pti Ii of d tach, d fHilts. After 6 days, significant larval mortality and r dU'cd growth of any surviving I( 'a was ob rv d clue to the high lev Is of Cry7Ar ins cticidal prot in :pr ssion re'lIlt d in th transform.cl brinja1 tissue. Among th nine events, VR13T-8 event was selected as the best cnt. in 1',; and 1'6 stag . Furth r, bi saf ly studi s are being carried out in the best s 'Ieel d nL
ruit- and Shoot-Borer Resistant Transgenic Brinjal
Al ng with Cry7A, a synth ti Cry7Aa3 g'o with full co I n-modification was aL 0 used to develop . r sistant brinjal. ran g ni's were developed by Agrobacterium-mediated plant transformation. Total R indepencl nt transformation events were regenerated from transform d Us of h 'pocot ,1 explants. peR and Southern blot nal e 'on mn d the pr ence of single opy ins rti n of the Cry7Aa3 g ne in plants of f ur inel pend nt tran formation v als. In addition, sin tie copy g a in ru n ~ r also confirm d by se!ITefTalion analy i of 1'1 seedlings. Expr ssion of Cry 7Aa3 transcripts wa
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CrylAc brinjal
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January-February 2016
CrylAa3 brinjal
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Transgenic Brinjal India is the second largest producer of brinjal with a total production of 11.89 million tonnes from 0.68 million ha area. but in terms of productivity (117.5 tonnes/ha). it lags behind many countries. The major bottleneck in increasing brinjal production is the brinjal fruit- and shoot-borer that results up to 65% yield loss. Presently. it is controlled mainly by massive application of insecticides which is often ineffective. Frequent and indiscriminate use of insecticides by farmers has also led to resistance development in the insects. increased costs of production. threatens human health and causes many environmental hazards. The lack of any natural source of resistance to FSB in al'l cultivated and conventionally cross-compatible species of Solanum is a major obstacle to developing FSB resistant cultivars. Using plant genetic engineering approach. we have developed FSB resistant transgenic eggplant lines by transferring different versions of cry genes (Cry lAc and Cry I Aa3) in cv. Kashi Taru, which are likely to be released in near future for commerci I cultivation. It is expected that Bt-brinjal would help farmers to effectively control FSB with a significant reduction in insecticide sprays. Furthermore, the different versions of cry genes (Cry I Ac and Cry I Aa3) could also be used for stacking into one variety. The trait stacked Bt transgenic plants have been proven e fective to prevent the emergence and build-up of insects resistant to Bt toxins. In India. considering the wide range of brinjal types (colo I', shape. size. and presence of spines on the calyx) and local preferences of a particular type. such cry genes could be br-ed to develop FSB resistant transgenic lines in different genetic backgrounds. Resource-limited farmers stand to benefit economically from the developed Bt brinjal lines by (i) High quality brinjal seeds available from public-sector sources at affordable prices. (ii) Reduced production losses from brinjal fruit- and shoot-borer. (iii) Increase in clean, marketable fruit yield. (iv) Reduced production costs due to less use of pesticide sprays. Considering the present and future status of the extensive use of insecticides to control FSB. host plant resistance via Cry I Ac-transformed plants may prOVide a first line of defence in an Integrated Pest Management (IPM) programme, prOVided transgenic food crops ar-e eventuaily approved and released for commercial cultivJtion in India.
con IrIDed by qRT-PCR and Ell 1A I' V aled Sigl1ifi 'anLly higher I >vels of Cry 7Aa3 pr ( iu in fresb lea es (3 .f.J 44.3 ng/g) and frui (2 ).5-::35.7 ng/g). Hjuh 1- v .I f expression of the Cry7Aa3 prot in r suIled in high F B larval m rlality in the hoot a w 11 ,- fruil tissues of the transformed brinjal lines. u h tran geni brinjal lin's posse. ing re istance to the ouJd r duce d pendency 00 pc ti ide use for FSB control, leading to a sal' r environill nt and brinjal fruits. hese transgenic plants are being tudied for bi safety,
Fruit-Borer Resistant Transgenic Tomato Tomato Fruit Borer (HeLicoverpa annigera) is ne of the m st d slru (i pests fe cting I mato pr duc .on. Improved inbr d line f tomato (Ka hi Vi h sh) was transformed b Agrobacterium-m diated plant transformation haVing binar vector pEin ill carrying a Cry7Ac g ne fused to a onsti utive C II '5 promoLer. The Agrobact ium wa co- uHivaLed \ ith pr -cultu ed cotyledon explants, and a lo te I 0 I' .5 Til transformants w r regeoeraL d. Th pr n of lh lran gene as confirmed b peR amplifi ati n. uth m anal si - shm d 16 i.ndependent transgenic ev nts were havine, a inrrl CO?" ry 7 c ene integr Lion. ELI analyse using a monoclonal antibody against ry 7Ac protein, Cry 1Ac
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r veal d th pr en e f Cry7Ac prot in and uantitativ UmaL on Iml d . i!,'TIilicant I v Is of Cry7Ac prot ill e pI' S ed in R lTan,'form d events. High I vels f expr ion of th in eti idal prot in r 'lilt d in a high mortalit rat of Lh larva f eding 0 fruits of tran form d LOmato plant. Homoz 'g us plan of Lh 8 e enls ( IT 1 Lo VRTT-8) of Cry7Ac transg nic tomato w re advanc d to T~ generati nand bj ted to event s Ie tion trial, In t bi as ay by f ding s ond-instar T B larva on Lh· ruits f Lransforme and non-Iran formed control plant: sh w d that the v nL had lhe high t mean mortalily and \ i ht loss aft r 9 days of ino 'ulation. Event s 1 '[j 11 trial as done and best ev n were furth r sturn d for biosaf ty. e
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Drought and Salt tress Tolerant Transgenic Tomato PI n frequ nlly nCQunter wal r-deficit stress dL! Lo drought, lei and saliniL at 'om stage of tn ir life. We ler d ficit stre 'trangly affe t' plants by altering it physiologi al functions, caus rcacti oxyg n sp -cies generation, ultimat I)' reducing the plant growth and fruit yi ld. Therefore, the capa"ity of plants 0 ithstand watcr~deficit slr s has assumed a gr 'at economic significance in ever chancrin cr climate. To impro e drought toleran e uf Lomato, AtDREB7AI CBF3 transcription tomato
Indian Horticulture
Transgenic Bt tomato Tomato is the most important vegetable among solanaceous crops and is largely cultivated by Indian farmers. The marketable yield of tomato fruits is affected mainly by tomato fruit borer. Huge amoUnt of insecticides are used by farmers to control tomato fruit borer. Along with increase in cost of production, use of insecticide is harmful to human health and environment. Like brinjal. no source of resistance is available in genotypes of tomato. Therefore, development of transgenic tomato providing resistance to tomato fruit borer is an environment friendly mode to control tomato fruit borer effectively. We have devleped transgenic tomato carrying Cry lAc gene in Kashi Vishesh cultivar. This cultivar can further be used in breeding programmes to transfer Cry lAc gene in other popular cultivars of tomato. Commercial cultivation of Bt tomato can save cost of production by less or no use of harmful insecticides. High yield of insect free and healthy tomato fruits with less cost of cultivation helps farmers to gain prosperity.
factor driven by water-deficit-induciblc promoter rd2.9A tomato evenLs show enhanced tolerance to drought and was transformed in tomato cV. H-8G (Kashi Vishesh). salt stresses. Selected events are being advanced to next Presence and integration of the tran.sgene were confinned generation for biosafety study. by PCR amplification and Southern blot analysis, respectively. Fifty days old T 2 plants of the 36 single Tolerant Transgenic Tomato using BcZAT12 Gene copy events were imposed to different drought stress To impart drought tolerance in tomato, another gene, lreaiments by withholding inigation for 7 days. 14 dal~' BcZAT12 driven by stress inducible lea 7 promoter was or 21 days. Over expression of rd29A::AtDREB1A/CBF3 used for AgrobacteTium-mediated transformation of tomato imparted a significant (cv. Kasha Vishesh). drought tolerance gain The presence of the in the transgenic tra nsgene wa .,,, tomato lines reilected confirmed by PCR IlH I.B by greater biomas~, amplification in ] 7 relative growth rate independent events and (RGR), and 12.6-23.00/1, transgcne integration higher relative water and copy number was content (RWC) observed by segregation associated with lower analysis. Mendelian membrane damage, inheritance was \~ lower chlorophyll aDd observed in 10 events. carolenoids reduction, The real time AtDREB lA tomato higher accumulation of expression of these 10 proline and soluble events were analysed ugars, and lower content of reactive oxygen species after 2 hours of heat exposure at ~2°C and different (ROS). The drought stressed AtDREB7A transgenic tomato water deficit (7, 14 and 21 days water withheld). Th plants revealed significantly higher specific activities of leaves of both types of stress plants were collected and key antioxidant enzymes namely, superoxide dismutas data were recorded for relative "..,tter conLent (RWC), (SOD), catalase (CAT), ascorbate peroxidase (APX), lectrolytic leakage, chlorophyll color index (Cel) glutathione reductase (GR), dehydro ascorbate reductase hydrogen perOxide level, MDA content and activity of HAR), and monodehydro ascorbate reductase (MDHAR). antioxidant enzymes (CAT, APX, POD, GR and SOD). The concentrations of non-enzymatic antioxidants ascorbic RWC of ZAT72 transgenjc hnes after drought anel heat acid and glutathione, and their reduced forms were higher stress were 17.0~-23.91"/LJ and 2.15-7.21% higherthall wild in the transgenic plants. The rd29A::AtDREB7A transgenic type (H-H6). There was 0.61-11.08% anel 6.cl3-12.74 u/o less tomato lines suppressed drought-il]duced oxidative stress ion leakage observed both in drought and heat stress in transgeniC lines. The by activating the 2.(i7-15.33 cV
