Symptoms of severe stunting, little leaf, yellowing and axillary shoot proliferation were recorded on chickpea plants in three northern states of India (New Delhi, Uttar Pradesh and Rajasthan) during January–March, 2017–2018 with disease incidences of 6–56%. Nested PCR assays with phytoplasma-specific universal primer pairs (P1/P7 and 3Far/3Rev) and 16S rRNA gene sequence analysis confirmed the association of ‘Candidatus Phytoplasma trifolii’ in the symptomatic chickpea variety Pusa Bheema at New Delhi. The in silico RFLP and phylogeny analyses of 16S rRNA sequence of the chickpea stunt (CpS) affected samples confirmed the association of 16SrVI-D subgroup phytoplasma. Similar phytoplasma strain (16SrVI-D subgroup) was detected in the leafhopper species, Hishimonus phycitis collected from symptomatic chickpea fields at New Delhi. Association of icosahedral virus particles of ~25–30 nm diameters were seen under electron microscope in CpS affected samples as well as in the aphids, Myzus persicae (Sulzer) collected from the symptomatic CpS samples at New Delhi. Both the plant and aphid samples were tested positive with the polyclonal antibodies (pAbs) of cucumber mosaic virus (CMV) in DAS-ELISA and ISEM tests. PCR assays using coat protein (CP) gene specific primers for CMV and the resultant sequence comparison also confirmed the association of CMV with the CpS affected symptomatic chickpea plants and M. persicae. The CpS affected samples collected from all the three states were further tested positive with chickpea chlorotic dwarf virus (chickpea chlorotic dwarf virus, CpCDV) CP gene specific primers in PCR assays. The 2.7 kb DNA sequence resulted from the rolling circle amplification (RCA) from the CpS affected chickpea samples of New Delhi revealed a highest sequence similarity of 99% with the CpCDV isolate from Pakistan. These results suggested a wider occurrence of CpCDV in chickpea crops in all the three states and a mixed infection of phytoplasma [‘Ca. P. trifolii’ (16SrVI-D subgroup)] and two plant viruses (CMV and CpCDV) associated with CpS disease at New Delhi.

2017年1月至3月，印度北部三个州（新德里，北方邦和拉贾斯坦邦）的鹰嘴豆植物上出现了严重的发育迟缓，叶片少，发黄和腋生芽的症状，发病率为6-56％。使用植物浆体特异性通用引物对（P1 / P7和3Far / 3Rev）和16S rRNA基因序列分析进行的巢式PCR分析证实了新德里有症状鹰嘴豆变种Pusa Bheema中的“ Candidatus Phytoplasma trifolii”的关联。鹰嘴豆特技（CpS）感染样品的16S rRNA序列的计算机RFLP和系统发育分析证实了16SrVI-D亚组植物质浆的相关性。在叶蝉物种Hismonus phycitis中检测到类似的质原体菌株（16SrVI-D亚组）收集自新德里有症状鹰嘴豆田。在电子显微镜下，在受CpS影响的样品以及从新德里有症状CpS样品中收集的蚜虫Myzus persicae（Sulzer）中观察到直径约25–30 nm的二十面体病毒颗粒的关联。在DAS-ELISA和ISEM测试中，用黄瓜花叶病毒（CMV）的多克隆抗体（pAbs）对植物和蚜虫样品进行了阳性测试。使用针对CMV的外壳蛋白（CP）基因特异性引物进行PCR分析并进行序列比较也证实了CMV与受CpS影响的有症状鹰嘴豆植物和桃蚜的相关性。从所有三个州收集的受CpS影响的样品进一步用鹰嘴豆绿藻矮化病毒（鹰嘴豆绿藻矮化病毒，CpCDV）CP基因特异性引物进行PCR检测呈阳性。来自新德里受CpS感染的鹰嘴豆样品的滚环扩增（RCA）产生的2.7 kb DNA序列显示，与来自巴基斯坦的CpCDV分离株的最高序列相似性为99％。这些结果表明CpCDV在鹰嘴豆作物中的所有三种状态更宽的发生和植原体的混合感染[”钙。P. trifolii'（16SrVI-D亚组）]和两种与新德里CpS病相关的植物病毒（CMV和CpCDV）。