Mushtaq Rakhshanda, Shahzad Khurram, Mansoor Shahid, Shah Zahid Hussain, Alsamadany Hameed, Mujtaba Tahir, Al-Zahrani Yahya, Alzahrani Hind A S, Ahmed Zaheer, Bashir Aftab
Agriculture Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan.
Department of Biotechnology, Pakistan Institute of Engineering and Applied Sciences, Nilore, Islamabad, Pakistan.
AoB Plants. 2018 Oct 16;10(6):ply067. doi: 10.1093/aobpla/ply067. eCollection 2018 Dec.
Cotton leaf curl virus (CLCuV) disease is one of the major limiting factors in cotton production, particularly in widely cultivated varieties that are susceptible to attack by this virus. Several approaches have been employed to explore putative resistance genes in another cotton species, . However, the exact mechanisms conferring disease resistance in cotton are still unknown. In the current study, we used various approaches to identify possible resistance genes against CLCuV infection. We report the identification and isolation of a set of genes involved in the resistance response to viral infestation. PCR products containing genomic DNA gave multiple amplifications with a single primer in most reactions, and 38 fragments were cloned from and . The sequences of cloned fragments belonged to various pathway genes and uncharacterized proteins. However, five amplified fragments (RM1, RM6, RM8, RM12 and RM31) showed similarity with R genes. Maximum homology (94 %) was observed with toll/interleukin receptor-like protein. BLAST search showed the homology of all resistance gene analogues (RGAs) with more than one chromosome, and multiple hits were observed on each chromosome for each RGA. Expression analysis through RT-PCR identified variable expression levels of the different RGAs in all tested genotypes. The expression level of RGAs differed between symptomatic and asymptomatic plants, with the exception of RGA 395, whose expression level was the same in both diseased and healthy plants. Knowledge of the interaction of these genes with various cotton pathogens could be utilized to improve the resistance of susceptible and other plant species.
棉花卷叶病毒(CLCuV)病是棉花生产中的主要限制因素之一,尤其是在广泛种植且易受该病毒侵袭的品种中。人们已采用多种方法在另一种棉花物种中探寻潜在的抗性基因。然而,棉花中赋予抗病性的确切机制仍不明确。在本研究中,我们运用多种方法来鉴定可能抵抗CLCuV感染的基因。我们报告了一组参与对病毒侵染抗性反应的基因的鉴定与分离。在大多数反应中,包含基因组DNA的PCR产物用单一引物进行了多次扩增,并且从[具体来源1]和[具体来源2]克隆了38个片段。克隆片段的序列属于各种途径基因和未表征的蛋白质。然而,五个扩增片段(RM1、RM6、RM8、RM12和RM31)与R基因显示出相似性。与toll/白细胞介素受体样蛋白的同源性最高(94%)。BLAST搜索显示所有抗性基因类似物(RGAs)与不止一条染色体具有同源性,并且每个RGA在每条染色体上都观察到多个匹配。通过RT-PCR进行的表达分析确定了所有测试基因型中不同RGAs的可变表达水平。有症状和无症状植物中RGAs的表达水平不同,但RGA 395除外,其在患病和健康植物中的表达水平相同。了解这些基因与各种棉花病原体的相互作用可用于提高易感[棉花品种名称]和其他植物物种的抗性。