Department of Agricultural Biotechnology, Anand Agricultural University, Anand, 388110, Gujarat, India.
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, 388110, Gujarat, India.
Phytochemistry. 2023 Sep;213:113776. doi: 10.1016/j.phytochem.2023.113776. Epub 2023 Jun 30.
Castor is industrially important non-edible oil seeds crop severely affected by soil borne pathogen Fusarium oxysporum f. sp. ricini which causes heavy economic losses among the castor growing states in India and worldwide. The development of Fusarium wilt resistant varieties in castor is also challenging because the genes identified for resistance are recessive in nature. Unlike transcriptomics and genomics, proteomics is always a method of choice for quick identification of novel proteins expressed during biological events. Therefore, comparative proteomic approach was employed for identification of proteins released in resistant genotype during Fusarium infection. Protein was extracted from inoculated 48-1 resistant and JI-35 susceptible genotype and subjected to 2D-gel electrophoresis coupled with RPLC-MS/MS. This analysis resulted in 18 unique peptides in resistant genotype and 8 unique peptides in susceptible genotype were identified through MASCOT search database. The real time expression study showed that 5 genes namely CCR 1, Germin like protein 5-1, RPP8, Laccase 4 and Chitinase like 6 was found highly up-regulated during Fusarium oxysporum infection. Furthermore, end point PCR analysis of c-DNA showed amplification of three genes namely Chitinase 6 like, RPP8 and β-glucanase exclusively in resistant genotype indicating that these genes may be involved in resistance phenomenon in castor. Up-regulation of CCR-1 and Laccase 4 involved in lignin biosynthesis provides mechanical strength and may help to prevent the entry of fungal mycelia and protein Germin like 5-1 helps to neutralized ROS by SOD activity. The clear role of these genes can be further confirmed through functional genomics for castor improvement and also for development of transgenic in different crops for wilt resistance.
蓖麻是一种重要的工业非食用油料作物，严重受到土壤病原菌尖孢镰刀菌（Fusarium oxysporum f. sp. ricini）的影响，这给印度和全球的蓖麻种植州造成了巨大的经济损失。在蓖麻中培育抗枯萎病品种也具有挑战性，因为鉴定出的抗性基因本质上是隐性的。与转录组学和基因组学不同，蛋白质组学一直是快速鉴定生物事件中表达的新蛋白质的首选方法。因此，采用比较蛋白质组学方法来鉴定抗尖孢镰刀菌感染的蓖麻基因型中释放的蛋白质。从接种的 48-1 抗性和 JI-35 敏感基因型中提取蛋白质，并进行 2D 凝胶电泳与 RPLC-MS/MS 结合。通过 Mascot 搜索数据库鉴定，在抗性基因型中鉴定出 18 个独特肽，在敏感基因型中鉴定出 8 个独特肽。实时表达研究表明，在尖孢镰刀菌感染过程中，有 5 个基因（即 CCR1、类 germin 蛋白 5-1、RPP8、漆酶 4 和几丁质酶 6）的表达显著上调。此外，c-DNA 的终点 PCR 分析显示，在抗性基因型中，仅扩增了三个基因，即几丁质酶 6 样、RPP8 和β-葡聚糖酶，表明这些基因可能参与了蓖麻的抗性现象。参与木质素生物合成的 CCR-1 和漆酶 4 的上调为机械强度提供了支持，可能有助于阻止真菌菌丝的进入，而类 germin 蛋白 5-1 通过 SOD 活性帮助中和 ROS。通过功能基因组学进一步证实这些基因的明确作用，可以促进蓖麻改良，也可以为不同作物的抗枯萎病转基因开发提供帮助。
