Institute of Plant Breeding and Biotechnology, MNS University of Agriculture, Multan, Pakistan.
Department of Biochemistry and Biotechnology, The Women University, Multan, Pakistan.
Mol Biol Rep. 2022 Jun;49(6):5521-5534. doi: 10.1007/s11033-022-07432-y. Epub 2022 May 12.
To supply high-quality cotton fibre for the textile industry, the development of long, strong and fine fibre cotton varieties is imperative. An interlinked approach was used to comprehend the role of fibre genes by analyzing interspecific progenies of cotton species. Wild Gossypium species and races are rich source of genetic polymorphism due to environmental dispersal and continuous natural selection. These genetic resources hold mass of outclass genes that can be used in cotton improvement breeding programs to exploit possible traits such as fibre quality, abiotic stress tolerance, and disease and insect resistance. Therefore, use of new molecular techniques such as genomics, transcriptomics and bioinformatics is very important to utilize the genetic potential of wild species in cotton improvement programs.
Interspecific lines and Gossypium species used in the study were grown at Central Cotton Research Institute (CCRI), Multan. After retrieving DNA sequence of the genes from NCBI, the primers for gene expression and full-length gene sequence were designed. Expression profiling of Expansin A4, BURP Domain protein RD22-like and E6-like fibre genes was performed through Real Time PCR. BLAST and DNA sequence alignment was conducted for sequence comparison of interspecific lines and Gossypium species. Different in silico analysis were used for characterization of fibre genes and identification of cis acting promoter elements in promoter region.
Variable expression of genes related to fibre development was observed at different stages. BLAST and DNA sequence alignment demonstrated resemblance of interspecific lines with G. hirsutum. In silico analysis on the sequence data also confirmed the role of Expansin A4, BURP Domain protein RD22-like and E6-like fibre genes in fibre development. Genetic engineering is also recommended by transferring E6-like, Expansin A4 and BURP Domain RD22-like genes in local cotton cultivars. Similarly, several stress tolerant and light responsive cis acting elements were identified through promotor analysis, which may contribute for fibre development in the breeding programs.
Expansin A4, BURP Domain RD22-like and E6-like have positive role in fibre development with variable expression at fiber length and strength associated stages.
为了向纺织业提供高质量的棉花纤维，开发长、强、细纤维的棉花品种势在必行。本研究采用种间杂种的方法，通过分析棉花种间杂种，来理解纤维基因的作用。由于环境扩散和持续的自然选择，野生棉种和棉种群体丰富，遗传多态性丰富。这些遗传资源蕴藏着大量优秀的基因，可以用于棉花改良育种计划，以利用纤维质量、非生物胁迫耐受性、疾病和虫害抗性等可能的特性。因此，利用基因组学、转录组学和生物信息学等新的分子技术，对于利用野生种的遗传潜力，在棉花改良计划中非常重要。
本研究中使用的种间系和棉种在中央棉花研究所（CCRI），木尔坦种植。从 NCBI 中检索到基因的 DNA 序列后，设计了基因表达和全长基因序列的引物。通过实时 PCR 对 Expansin A4、BURP 结构域蛋白 RD22 样和 E6 样纤维基因的表达谱进行了分析。对种间系和棉种的序列进行了 BLAST 和 DNA 序列比对，以比较序列。利用不同的计算机分析方法对纤维基因进行了特征描述，并鉴定了启动子区域中的顺式作用启动子元件。
在不同的发育阶段，观察到与纤维发育相关的基因的表达存在差异。BLAST 和 DNA 序列比对表明，种间系与 G. hirsutum 相似。对序列数据的计算机分析也证实了 Expansin A4、BURP 结构域 RD22 样和 E6 样纤维基因在纤维发育中的作用。遗传工程也建议通过转移 E6 样、Expansin A4 和 BURP 结构域 RD22 样基因到当地棉花品种中。同样，通过启动子分析鉴定了几个应激耐受和光响应的顺式作用元件，这可能有助于在育种计划中进行纤维发育。
Expansin A4、BURP 结构域 RD22 样和 E6 样在纤维发育中起积极作用，在纤维长度和强度相关阶段表现出不同的表达。
