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首页> 外文期刊>Pakistan journal of botany >AGROBACTERIUM MEDIATED TRANSFORMATION OF BRASSICA JUNCEA (L.) CZERN. WITH CHITINASE GENE CONFERRING RESISTANCE AGAINST FUNGAL INFECTIONS
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AGROBACTERIUM MEDIATED TRANSFORMATION OF BRASSICA JUNCEA (L.) CZERN. WITH CHITINASE GENE CONFERRING RESISTANCE AGAINST FUNGAL INFECTIONS

机译：农杆菌介导的甘蓝型juncea（L.）Czern的转化。用几丁质酶基因赋予抗真菌感染抗性

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摘要

Brassica juncea (Czern & Coss., L.) is an important oilseed crop. Since it is attacked by several bacterial and fungaldiseases, therefore, we developed an easy and simple protocol for the regeneration and transformation of B. juncea varietyRAYA ANMOL to give rise to transgenic plants conferring resistance against various fungal diseases. The transformationwas carried out using Agrobacterium with Chitinase gene. This gene was isolated from Streptomyces griseus HUT6037. Weused two types of explants for transformation i.e. hypocotyls and cotyledons. Only hypocotyls explants showed good resultsregarding callus initiation. Different hormonal concentrations were applied i.e. BAP 2, 4 and 6 mgL-1 and NAA 0.1, 0.2 and0.3 mgL-1. However, high transformation efficiency was observed by supplementing the medium with combination of 2mgL-1 BAP and 0.2 mgL-1 for initiation of callus. Similarly 10 mgL-1 kanamycin and 200 mgL-1 cefotaxime also provedsuccessful for the selection of transformed callus. In order to confirm the presence of transgenic callus Polymerase chainreaction was performed using specific primers for Chitinase gene.
机译：Brassica Juncea（Czern＆Coss，L.）是一个重要的油籽作物。因此，由于它受到几种细菌和真菌的攻击，因此我们开发了一种简单而简单的协议，用于B. juncea varietyraya Anmol的再生和转化，从而产生赋予针对各种真菌疾病的转基因植物。使用含量蛋白酶基因的农杆菌进行转化。该基因与链霉菌族Hut6037分离出来的。湿润两种转化的外植体I.对胚乳和子叶。只有次髓基底植物才显示出良好的竞争愈伤组织启动。施用不同的激素浓度，即BAP 2,4和6 MgL-1和NAA 0.1,0.2和0.3mgL-1。然而，通过补充具有2mgL-1 BAP的组合和0.2MgL-1的培养基来观察到高转化效率，用于启动愈伤组织。同样地，10 mgl-1 kanamycin和200 mgl-1 cefotaxime也证明了选择转化的愈伤组织。为了确认转基因愈伤组织聚合酶的存在，使用特异性引物进行逐蛋白酶基因进行。

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