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整合代谢转录组学揭示小麦QTL-Fhb2中赤霉病候选抗性基因

Integrated Metabolo-Transcriptomics Reveals Fusarium Head Blight Candidate Resistance Genes in Wheat QTL-Fhb2.

作者信息

Dhokane Dhananjay, Karre Shailesh, Kushalappa Ajjamada C, McCartney Curt

机构信息

Department of Plant Science, Macdonald Campus, McGill University, 21,111 Lakeshore Road, Sainte-Anne-de-Bellevue, Quebec, H9X 3V9, Canada.

Agriculture and Agri-Food Canada, 195 Dafoe Road, Winnipeg, Manitoba, R3T 2M9, Canada.

出版信息

PLoS One. 2016 May 27;11(5):e0155851. doi: 10.1371/journal.pone.0155851. eCollection 2016.

Abstract

BACKGROUND

Fusarium head blight (FHB) caused by Fusarium graminearum not only causes severe losses in yield, but also reduces quality of wheat grain by accumulating mycotoxins. Breeding for host plant resistance is considered as the best strategy to manage FHB. Resistance in wheat to FHB is quantitative in nature, involving cumulative effects of many genes governing resistance. The poor understanding of genetics and lack of precise phenotyping has hindered the development of FHB resistant cultivars. Though more than 100 QTLs imparting FHB resistance have been reported, none discovered the specific genes localized within the QTL region, nor the underlying mechanisms of resistance.

FINDINGS

In our study recombinant inbred lines (RILs) carrying resistant (R-RIL) and susceptible (S-RIL) alleles of QTL-Fhb2 were subjected to metabolome and transcriptome profiling to discover the candidate genes. Metabolome profiling detected a higher abundance of metabolites belonging to phenylpropanoid, lignin, glycerophospholipid, flavonoid, fatty acid, and terpenoid biosynthetic pathways in R-RIL than in S-RIL. Transcriptome analysis revealed up-regulation of several receptor kinases, transcription factors, signaling, mycotoxin detoxification and resistance related genes. The dissection of QTL-Fhb2 using flanking marker sequences, integrating metabolomic and transcriptomic datasets, identified 4-Coumarate: CoA ligase (4CL), callose synthase (CS), basic Helix Loop Helix (bHLH041) transcription factor, glutathione S-transferase (GST), ABC transporter-4 (ABC4) and cinnamyl alcohol dehydrogenase (CAD) as putative resistance genes localized within the QTL-Fhb2 region.

CONCLUSION

Some of the identified genes within the QTL region are associated with structural resistance through cell wall reinforcement, reducing the spread of pathogen through rachis within a spike and few other genes that detoxify DON, the virulence factor, thus eventually reducing disease severity. In conclusion, we report that the wheat resistance QTL-Fhb2 is associated with high rachis resistance through additive resistance effects of genes, based on cell wall enforcement and detoxification of DON. Following further functional characterization and validation, these resistance genes can be used to replace the genes in susceptible commercial cultivars, if nonfunctional, based on genome editing to improve FHB resistance.

摘要

背景

禾谷镰刀菌引起的小麦赤霉病不仅会导致严重的产量损失,还会通过积累霉菌毒素降低小麦籽粒品质。培育寄主植物抗性被认为是防治小麦赤霉病的最佳策略。小麦对赤霉病的抗性本质上是数量性状,涉及许多抗性相关基因的累积效应。对遗传学的了解不足以及缺乏精确的表型分析阻碍了抗赤霉病品种的培育。尽管已经报道了100多个赋予赤霉病抗性的QTL,但没有一个发现位于QTL区域内的特定基因,也没有发现抗性的潜在机制。

研究结果

在我们的研究中,对携带QTL-Fhb2抗性(R-RIL)和感病(S-RIL)等位基因的重组自交系进行了代谢组和转录组分析,以发现候选基因。代谢组分析检测到,与S-RIL相比,R-RIL中属于苯丙烷类、木质素、甘油磷脂、类黄酮、脂肪酸和萜类生物合成途径的代谢物丰度更高。转录组分析揭示了几种受体激酶、转录因子、信号传导、霉菌毒素解毒和抗性相关基因的上调。利用侧翼标记序列对QTL-Fhb2进行剖析,整合代谢组学和转录组学数据集,确定4-香豆酸:辅酶A连接酶(4CL)、胼胝质合酶(CS)、碱性螺旋环螺旋(bHLH041)转录因子、谷胱甘肽S-转移酶(GST)、ABC转运蛋白4(ABC4)和肉桂醇脱氢酶(CAD)为位于QTL-Fhb2区域内的假定抗性基因。

结论

QTL区域内一些已鉴定基因通过增强细胞壁与结构抗性相关,减少病原菌在穗轴内通过穗轴的传播,还有一些基因对致病因子脱氧雪腐镰刀菌烯醇(DON)进行解毒,从而最终降低病害严重程度。总之,我们报道小麦抗性QTL-Fhb2通过基于细胞壁强化和DON解毒的基因加性抗性效应与高穗轴抗性相关。经过进一步的功能表征和验证后,如果易感商业品种中的基因无功能,这些抗性基因可用于基于基因组编辑来替代这些基因,以提高对小麦赤霉病的抗性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0e08/4883744/a5a1dd9402ec/pone.0155851.g001.jpg

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