Upasani Medha L, Gurjar Gayatri S, Kadoo Narendra Y, Gupta Vidya S
Biochemical Sciences Division, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pashan, Pune 411008, India.
PLoS One. 2016 May 26;11(5):e0156490. doi: 10.1371/journal.pone.0156490. eCollection 2016.
Fusarium wilt caused by Fusarium oxysporum f.sp. ciceri (Foc) is a constant threat to chickpea productivity in several parts of the world. Understanding the molecular basis of chickpea-Foc interaction is necessary to improve chickpea resistance to Foc and thereby the productivity of chickpea. We transformed Foc race 2 using green fluorescent protein (GFP) gene and used it to characterize pathogen progression and colonization in wilt-susceptible (JG62) and wilt-resistant (Digvijay) chickpea cultivars using confocal microscopy. We also employed quantitative PCR (qPCR) to estimate the pathogen load and progression across various tissues of both the chickpea cultivars during the course of the disease. Additionally, the expression of several candidate pathogen virulence genes was analyzed using quantitative reverse transcriptase PCR (qRT-PCR), which showed their characteristic expression in wilt-susceptible and resistant chickpea cultivars. Our results suggest that the pathogen colonizes the susceptible cultivar defeating its defense; however, albeit its entry in the resistant plant, further proliferation is severely restricted providing an evidence of efficient defense mechanism in the resistant chickpea cultivar.
由尖孢镰刀菌鹰嘴豆专化型(Foc)引起的枯萎病对世界上多个地区的鹰嘴豆产量构成持续威胁。了解鹰嘴豆与Foc相互作用的分子基础对于提高鹰嘴豆对Foc的抗性从而提高鹰嘴豆产量至关重要。我们用绿色荧光蛋白(GFP)基因转化了Foc小种2,并利用共聚焦显微镜对其在感病(JG62)和抗病(Digvijay)鹰嘴豆品种中的病原菌进展和定殖情况进行了表征。我们还采用定量PCR(qPCR)来估计在病程中两个鹰嘴豆品种各组织中的病原菌载量和进展情况。此外,利用定量逆转录PCR(qRT-PCR)分析了几个候选病原菌毒力基因的表达情况,结果显示它们在感病和抗病鹰嘴豆品种中具有特征性表达。我们的结果表明,病原菌定殖于感病品种并破坏其防御;然而,尽管它进入了抗病植株,但进一步增殖受到严重限制,这证明了抗病鹰嘴豆品种中存在有效的防御机制。