Generation of insect-resistant, transgenic crop plants by expression of the insecticidal crystal protein (ICP) gene of Bacillus thuringiensis (Bt) is a standard crop improvement approach. In such cases, adequate expression of the most appropriate ICP against the target insect pest of the crop species is desirable. It is also considered advantageous to generate Bt-transgenics with multiple toxin systems to control rapid development of pest resistance to the ICP. Larvae of yellow stem borer (YSB), Scirpophaga incertulas, a major lepidopteran insect pest of rice, cause massive losses of rice yield. Studies on insect feeding and on the binding properties of ICP to brush border membrane receptors in the midgut of YSB larvae revealed that cryIAb and cryIAc are two individually suitable candidate genes for developing YSB-resistant rice. Programs were undertaken to develop Bt-transgenic rice with these ICP genes independently in a single cultivar. A cryIAc gene was reconstructed and placed under control of the maize ubiquitin 1 promoter, along with the first intron of the maize ubiquitin 1 gene, and the nos terminator. The gene construct was delivered to embryogenic calli of IR64, an elite indica rice cultivar, using the particle bombardment method. Six highly expressive independent transgenic ICP lines were identified. Molecular analyses and insect-feeding assays of two such lines revealed that the transferred synthetic cryIAc gene was expressed stably in the T 2 generation of these lines and that the transgenic rice plants were highly toxic to YSB larvae and lessened the damage caused by their feeding.Technologies of plant cell biology and molecular biology have attracted much attention because they provide a powerful and novel means to supplement as well as complement the traditional methods of crop improvement. The transgenic approach of plant genetic engineering provides access to an unlimited gene pool for the transfer of desirable genes between any two species of interest, irrespective of their evolutionary or taxonomic relation. The successful generation of transgenic insectresistant crop plants by transferring the insecticidal crystal protein (ICP) gene of Bacillus thuringiensis (Bt; refs. 1-8), a Gram-positive, spore-forming bacterium, is one of the major breakthroughs in crop science that the new genetics has provided.indica rice is one of the most important food crops (9) of the world. There are half a dozen different species of lepidopteran insects whose larvae bore into the tissue of rice plants, and the resultant damage causes massive yield losses (10). Yellow stem borer (YSB), Scirpophaga incertulas, is most certainly the major contributor to these losses. The first-stage YSB larvae cut small holes in the stem of the paddy plant and enter the plant tissue, remaining there for the rest of their lives as larvae and pupae. The larvae feed voraciously inside the stem, providing themselves complete protection against chemical pesticides. There are no known resistance genes in rice (11) that can be rea...

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Three genotypes of chickpea ICCV-1, ICCV-6 and a Desi (local) variety were tested for plant regeneration through multiple shoot production. The embryo axis was removed from mature seeds, the root meristem and the shoot apex were discarded. These explants were cultured on medium containing MS macro salts, 4X MS micro salts, I35 vitamins, 3.0 mg/1 BAP, 0.004 mg/1 NAA, 3% (w/v) sucrose and incubated at 26(0)C. The explants were transformed withAgrobacterium tumefaciens strain LBA4404 with binary vector pBI121 containing theuidA andnptIl genes. Multiple shoots were repeatedly selected with kanamycin. The selected kanamycin resistant shoots were rooted on MS medium supplemented with 0.05 mg/1 113A. The presumptive transformants histochemically stained positive for GUS. Additionally, nptll assay confirmed the expression ofnptII in kanamycin resistant plants. Transgenic plants were transferred to soil and grown in the green house.

Using a biolistic particle delivery system, we have developed an efficient protocol for the generation of stable genetic transformants in jute (Corchorus capsularis var. JRC 321). The apical, meristematic region of a germinating seedling was used as the explant. Multiple shoots were generated from each explant using a medium supplemented with benzylaminopurine (0.2 mg/l) and indole-3-butyric acid (0.5 mg/l). Cotransformation was carried out with the bialaphos resistance gene bar and the rolC gene of Agrobacterium rhizogenes. The positive transformants containing the bar gene grew in growth medium containing 2 mg/l bialaphos. Southern, Northern, polymerase chain reaction (PCR) and reverse transcriptase-PCR analyses provided evidence of gene integration into the genomic DNA of jute. Transgenic plants were resistant to bialaphos. One plant had an altered growth pattern, most likely due to the expression of the transferred rolC gene. The T O transformants showed a stable inheritance of the trans gene to their progenies.

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