A detached petal disc assay and virus-induced gene silencing facilitate the study of Botrytis cinerea resistance in rose flowers
A detached petal disc assay and virus-induced gene silencing facilitate the study of Botrytis cinerea resistance in rose flowers
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分离花瓣盘测定和病毒诱导的基因沉默促进了月季花灰霉病抗性的研究
DOI：
10.1038/s41438-019-0219-2
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发表时间：
2019-12-01
影响因子：
8.7
通讯作者：
Zhang, Zhao
中科院分区：
文献类型：
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作者：
Cao, Xiaoqian;Yan, Huijun;Zhang, Zhao
鲜切玫瑰（Rosa hybrida）是世界上最重要的观赏作物之一，年贸易额达数十亿美元。灰霉病是危害切花月季最严重的一种真菌病害，造成了月季采后的严重损失。在这项研究中，我们优化了人工B的分离花瓣圆盘法（DPDA）。灰霉病接种和量化玫瑰花瓣中的疾病症状。此外，作为鉴定玫瑰基因参与B。为了研究灰霉病抗性基因B的功能，我们设计了一种病毒诱导的基因沉默（VIGS）方法。玫瑰花瓣中的灰霉病抗性基因。我们使用RhPR10。1作为沉默效率的报告者，发现玫瑰品种'Samantha'在RhPR10中表现出最大的降低。1在供试品种中的表达。确定茉莉酸和乙烯是否是B所必需的。在玫瑰花瓣中的灰霉病抗性中，我们使用VIGS沉默RhLOX 5和RhEIN 3（分别编码茉莉酸生物合成途径蛋白和乙烯调节蛋白）的表达，并发现花瓣对B的敏感性。灰叶受到影响。最后是B的VIGS屏幕。cinerea诱导的rose转录因子证明了该方法对于B中基因功能的高通量鉴定的潜在益处。灰霉病抗性总的来说，我们的数据表明，DPDA和VIGS的组合是一个可靠的和高通量的方法研究B。月季抗灰霉病。
Fresh-cut roses (Rosa hybrida) are one of the most important ornamental crops worldwide, with annual trade in the billions of dollars. Gray mold disease caused by the pathogen Botrytis cinerea is the most serious fungal threat to cut roses, causing extensive postharvest losses. In this study, we optimized a detached petal disc assay (DPDA) for artificial B. cinerea inoculation and quantification of disease symptoms in rose petals. Furthermore, as the identification of rose genes involved in B. cinerea resistance could provide useful genetic and genomic resources, we devised a virus-induced gene silencing (VIGS) procedure for the functional analysis of B. cinerea resistance genes in rose petals. We used RhPR10. 1 as a reporter of silencing efficiency and found that the rose cultivar ‘Samantha’showed the greatest decrease in RhPR10. 1 expression among the cultivars tested. To determine whether jasmonic acid and ethylene are required for B. cinerea resistance in rose petals, we used VIGS to silence the expression of RhLOX5 and RhEIN3 (encoding a jasmonic acid biosynthesis pathway protein and an ethylene regulatory protein, respectively) and found that petal susceptibility to B. cinerea was affected. Finally, a VIGS screen of B. cinerea-induced rose transcription factors demonstrated the potential benefits of this method for the high-throughput identification of gene function in B. cinerea resistance. Collectively, our data show that the combination of the DPDA and VIGS is a reliable and high-throughput method for studying B. cinerea resistance in rose.