Data_Sheet_1_Functional Verification of Two Genes Related to Stripe Rust Resistance in the Wheat-Leymus mollis Introgression Line M8664-3.pdf

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most widespread and destructive fungal diseases of wheat worldwide. The cultivation and growth of resistant wheat varieties are the most economical, effective, and environmental friendly methods to control stripe rust. Therefore, it is necessary to use new resistance genes to breed resistant wheat varieties. A single dominant gene temporarily designated as YrM8664-3, from a wheat-Leymus mollis introgression line M8664-3 highly resistant to Chinese predominant Pst races, is a potentially valuable source of stripe rust resistance for breeding. Herein, based on previous YrM8664-3 chromosome location results (bin 4AL13-0.59-0.66 close to 4AL12-0.43-0.59) and expression change information of candidate genes and bioinformatics analysis, several candidate genes with significantly different expression changes were then selected and verified by virus-induced gene silencing (VIGS). Two of the candidate genes temporarily designated as TaFBN [containing plastid lipid-associated proteins (PAP)_fibrillin domain in its protein] and Ta_Pes_BRCT [containing Pescadillo and breast cancer tumour suppressor protein C-terminus (BRCT) domain in its protein], produced the most significant resistance changes in the wheat-Pst interaction system after silencing. These two genes were further verified by Agrobacterium-mediated wheat genetic transformation technology. According to the identification of disease resistance, the resistance function of the candidate gene TaFBN was further verified. Then, the expression of TaFBN under hormone treatment indicated that TaFBN may be related to the salicylic acid (SA) and abscisic acid (ABA) signaling pathways. Combined with the expression of TaFBN in response to environmental stress stimulation, it can be reasonably speculated that TaFBN plays an important role in the resistance of wheat to Pst and is involved in abiotic stress pathways.

条纹锈病，由条形黑粉菌（Puccinia striiformis f. sp. tritici，简称Pst）引起，是全球范围内小麦最为普遍且破坏力极强的真菌病害之一。培育和种植抗病小麦品种，作为控制条纹锈病的最为经济、有效且环保的方法，显得尤为必要。因此，利用新的抗性基因培育抗病小麦品种显得尤为迫切。一种暂时命名为YrM8664-3的单显性基因，源自小麦与Leymus mollis的杂交线M8664-3，该杂交线对在中国普遍存在的Pst菌株表现出高度抗性，成为培育条纹锈病抗性的潜在宝贵资源。在本研究中，基于先前YrM8664-3染色体定位结果（位于4AL13-0.59-0.66附近，靠近4AL12-0.43-0.59）以及候选基因表达变化信息与生物信息学分析，我们选取并验证了数个表达变化显著不同的候选基因，通过病毒诱导的基因沉默（VIGS）技术进行了验证。其中两个候选基因，暂命名为TaFBN（其蛋白包含质体脂质相关蛋白（PAP）纤连蛋白结构域）和Ta_Pes_BRCT（其蛋白包含Pescadillo和乳腺癌抑制蛋白C端（BRCT）结构域），在沉默处理后，对小麦-Pst相互作用系统中的抗性变化产生了最为显著的影响。这两个基因随后通过农杆菌介导的小麦遗传转化技术进行了进一步验证。根据疾病抗性的鉴定，候选基因TaFBN的抗性功能得到了进一步的验证。随后，对TaFBN在激素处理下的表达研究显示，TaFBN可能与水杨酸（SA）和脱落酸（ABA）的信号通路相关。结合TaFBN对环境应激刺激的响应表达，可以合理推测，TaFBN在小麦对Pst的抗性中起着至关重要的作用，并参与了非生物胁迫途径。