Rabha Mihir, Sharma Shaswati, Acharjee Sumita, Sarmah Bidyut Kumar
Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat, Assam 785013 India.
DBT-AAU Centre, Assam Agricultural University, Jorhat, Assam 785013 India.
3 Biotech. 2017 Oct;7(5):303. doi: 10.1007/s13205-017-0935-y. Epub 2017 Sep 9.
We have identified both crystalliferous and acrystalliferous Bt isolates from the Assam soil of North East India for the first time. A total of 301 type colonies were selected based on their appearance and colony morphology. Out of these colonies, 42 isolates had characteristics similar to Bt isolates on MYP (Mannitol Egg Yolk Polymyxin) agar base medium. The ERIC-PCR and 16S rDNA analyses confirmed that 42 isolates are . Phase contrast microscopy showed that 37 isolates produced crystal endospore during the sporulation phase and 5 acrystalliferous isolates were also found. Amplification of gene was carried out using general Cry primers along with one gene specific primer. Out of 42 isolates, 50% of the isolates showed presence of gene followed by (40.47) and (40.47). Moreover, 21.42% of isolates showed the presence of more than one genes. We also screened these isolates for the possibility of having new Bt genes using universal primer and found two strains having a new type of gene with 82 and 85% similarities with the available gene sequences. Thus, these new types of Bt gene could be useful for Bt-based bioformulations and generation of transgenic plants.
我们首次从印度东北部阿萨姆邦的土壤中鉴定出了产晶体和不产晶体的苏云金芽孢杆菌(Bt)分离株。基于其外观和菌落形态,共挑选了301个典型菌落。在这些菌落中,有42个分离株在甘露醇卵黄多粘菌素(MYP)琼脂基础培养基上具有与Bt分离株相似的特征。ERIC-PCR和16S rDNA分析证实这42个分离株为……相差显微镜显示,37个分离株在芽孢形成阶段产生了晶体芽孢,同时也发现了5个不产晶体的分离株。使用通用的Cry引物以及一个cry基因特异性引物对cry基因进行扩增。在42个分离株中,50%的分离株显示存在cry基因,其次是cry……(40.47)和cry……(40.47)。此外,21.42%的分离株显示存在不止一种cry基因。我们还使用通用引物筛选了这些分离株中存在新Bt基因的可能性,发现有两个菌株具有一种新型的cry基因,与现有cry基因序列的相似性分别为82%和85%。因此,这些新型的Bt基因可能对基于Bt的生物制剂和转基因植物的培育有用。