海南文心兰切花品种病毒病的分子鉴定  

Molecular identification of viral diseases of Oncidium hybridum cutting flower in Hainan province

作　　者：闫钟荣 谢承智 宋希强[1,2] 王洪星[1,2] YAN Zhongrong;XIE Chengzhi;SONG Xiqiang;WANG Hongxing(Key Laboratory of Biology of Tropical Flower Resources of Hainan Province,Hainan University,Haikou 570228,Hainan,China;College of Forestry,Hainan University,Haikou 570228,Hainan,China)

机构地区：[1]海南大学海南省热带特色花木资源生物学重点实验室,海南海口570228 [2]海南大学林学院,海南海口570228

出　　处：《中南林业科技大学学报》2022年第11期53-62,共10页Journal of Central South University of Forestry & Technology

基　　金：海南省重点科技计划项目(ZDYF2020099);海南省自然科学基金项目(320MS007);海南省高等学校科学研究课题(Hnky2020-14);海南省重大科技专项(zdkj201815)。

摘　　要：【目的】文心兰切花产业占我国70%的市场,已经成为海南省重要的热带花卉支柱产业之一。文心兰生长过程中易受病毒的侵染,严重影响其植株生长、叶片发育、花形花色,同时降低其观赏价值,开展海南文心兰切花品种叶片病毒病的系统性分子鉴定有助于病害防治。【方法】本研究以海口、文昌、乐东、三亚、五指山、保亭、琼海、儋州8个主产区县市采集的12个品种共178份文心兰疑似病毒感染样品为试验材料,通过RT-PCR及小RNA(sRNA)深度测序技术进行病原分子鉴定,克隆其CP基因并进行生物信息学分析。【结果】1)海南文心兰疑似病毒感染样品叶片呈现褪绿状斑或条纹、黄化雾状斑、皱缩、黑色坏色斑等症状,且不同品种间存在差异,主要侵染病毒为建兰花叶病毒(Cymbidium mosaic virus, CymMV)和齿兰环斑病毒(Odontoglossum ringspot virus, ORSV);2)建兰花叶病毒海南分离物CymMV-HN和齿兰环斑病毒海南分离物ORSV-HN的CP基因核酸序列长度分别为672和477 bp,编码223和158个氨基酸,与已报道的病毒CP基因序列具有较高的同源性,表明在进化过程中具有较高的保守性;3)s RNA深度测序共获得26 976 941条raw reads,过滤后得到26 039 391条clean reads,拼接后获得1 256个Contigs,在Nr等数据库中共同注释到66个contigs,在CymMV和ORSV基因组数据库中分别注释到56和39个contigs,表明海南文心兰病毒病主要由CymMV和ORSV侵染而导致。【结论】CymMV和ORSV是海南文心兰感病的主要病毒类型。【Objective】Oncidium hybridum is an important ornamental plant species in Hainan province,which is sensitively infected with viruses during the growth process which seriously affects plant growth,leaf development,flower shape and color,and reduces ornamental value.However,the systematic molecular identification of viral diseases of Oncidium hybridum in Hainan province has not been reported.【Method】Using the Oncidium hybridum samples from 8 major producing areas in Hainan province with suspected viral infection as the experimental materials,the virus was detected and identified by reverse transcription polymerase chain reaction(RTPCR)and small RNA deep sequencing techniques.Meanwhile,the CP(Coat protein)gene was cloned and analyzed by bioinformatics.【Result】The infected leaves of Oncidium hybridum in Hainan province showed symptoms such as chlorotic spots or stripes,yellow mist spots,shrinkage and black spots,and there were significant differences among different varieties.The main infected viruses were Cymbidium mosaic virus(CymMV)and Odontoglossum ringspot virus(ORSV).2)The nucleotide sequences of the CP genes of CymMV-HN and ORSV-HN were 672 and 477 bp,encoding 223 and 158 amino acids which had a high homology with the existing viral CP gene sequence,indicating that there was high conservation during evolution.3)A total of 26976941 raw reads were obtained by small RNA deep sequencing,and 26039391 clean reads were obtained after filtering.The 1256 contigs were obtained after splicing,66 contigs were annotated in databases such as NR and so on,and 56 and 39 contigs were annotated in CymMV and ORSV genome databases,indicating that CymMV and ORSV were the main viruses of Oncidium hybridum virus diseases in Hainan province.【Conclusion】CymMV and ORSV are the main viruses infecting Oncidium hybridum in Hainan province.

关 键 词：海南文心兰 病毒 RT-PCR CP基因 sRNA深度测序 

分 类 号：S718.85[农业科学—林学]