玉米矮花叶病毒分离物基因组克隆及多样性分析
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摘要
玉米是重要的粮食和经济作物,在国民经济中占有重要地位。玉米矮花叶病毒(Sugercan mosaic virus,SCMV)广泛分布在世界各玉米主产区,严重威胁玉米的安全生产。系统研究SCMV的发生危害、遗传结构与进化机制,对病毒病的防治具有重要意义。从山西省4个玉米主产区采集122个玉米叶片样品(2015年62份,2016年60份)。经RT-PCR检测证实36个样品(2015年20个样品,2016年16个样品)为SCMV阳性,且广泛分布于山西省玉米各个产区。从这些阳性样品中分离、测序、克隆得到了一个新的SCMV分离物,该分离物(包括5'-UTR和3'-UTR端)基因组全长9 539 bp,编码3 063个氨基酸。该分离物与26个SCMV分离物(NCBI)进行一致率、系统发育分析表明SCMV存在较大遗传变异,27个分离物被划分为3个与地理位置无明显相关性的不同进化群体。选择压力分析表明,负向选择可能是SCMV遗传变异的原因之一。对变异位点统计分析发现,该结果将为评估SCMV在中国的流行病学特征奠定基础,有助于SCMV的长期可持续防控策略的制定。
Maize is an important food and economic crop,which plays an important role in the national economy.Sugercan mosaic virus widely distributed in the main maize producing areas in the world,and it is a great threat for the maize production. For controling SCMV,it is necessary to systematically study the damage,genetic structure and evolution mechanism. This study collected 122 maize leaf samples( 62 samples in 2015 and 60 samples in 2016) from four maize producing areas in Shanxi province. By RT-PCR,36 samples( 20 in 2015 and 16 in 2016) were proved to be positive for SCMV. One new SCMV strain was isolated and sequenced from these positive samples. It was found that the genome of this isolate was 9 539 bp including 5'-UTR and 3'-UTR region and encoded 3 063 aa. The identity and phylogenetic analysis with 26 isolates reported online NCBI were carried out,it was found the 27 isolates were divided into 3 different evolutionary populations based on the whole-genome. The results of this study provided a basis for assessing the epidemiological characteristics of SCMV in China and contribute to the formulation of long-term sustainable prevention and control strategies for SCMV.
Maize is an important food and economic crop,which plays an important role in the national economy.Sugercan mosaic virus widely distributed in the main maize producing areas in the world,and it is a great threat for the maize production. For controling SCMV,it is necessary to systematically study the damage,genetic structure and evolution mechanism. This study collected 122 maize leaf samples( 62 samples in 2015 and 60 samples in 2016) from four maize producing areas in Shanxi province. By RT-PCR,36 samples( 20 in 2015 and 16 in 2016) were proved to be positive for SCMV. One new SCMV strain was isolated and sequenced from these positive samples. It was found that the genome of this isolate was 9 539 bp including 5'-UTR and 3'-UTR region and encoded 3 063 aa. The identity and phylogenetic analysis with 26 isolates reported online NCBI were carried out,it was found the 27 isolates were divided into 3 different evolutionary populations based on the whole-genome. The results of this study provided a basis for assessing the epidemiological characteristics of SCMV in China and contribute to the formulation of long-term sustainable prevention and control strategies for SCMV.
引文
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[21]吕庆雪,高嵩,何欢,等.新型玉米种子生产技术表达载体的构建及遗传转化[J].基因组学与应用生物学,2017(6):2497-2501.Lv Q X, Gao S, He H, et al.. Construction and genetic transformation of expression vector in new-type maize seeds production technology[J]. Genomics Appl. Biol.,2017(6):2497-2501.
[2]张卫建.对我国玉米绿色增产增效栽培技术的探讨:增密减氮[J].作物杂志,2015(4):1-4.Zhang W J. The discuss on the cultivation approach to green improvement of maize yield and N use efficiency in China:Dense planting with Less N fertilizer[J]. Crops,2015(4):1-4.
[3]柴宗文,王克如,郭银巧,等.玉米机械粒收质量现状及其与含水率的关系[J].中国农业科学,2017,50(11):2036-2043.Chai Z W,Wang K R,Guo Y Q,et al.. Current status of maize mechanical grain harvesting and its relationship with grain moisture content[J]. Sci. Agric. Sin.,2017,50(11):2036-2043.
[4]郑向阳,栗建枝,王国平,等.山西省玉米不同生态区玉米产量性状差异化分析[J].中国农学通报,2015,31:68-74.Zheng X Y,Li J Z,Wang G P,et al.. The yield analysis of maize in different ecological area in Shanxi[J]. Chin. Agric.Sci. Bull.,2015,31:68-74.
[5]Xie X, Chen W,Fu Q,et al.. Molecular variability and distribution of Sugarcane mosaic virus in Shanxi,China[J].PLoS ONE,2016,11(3):e0151549.
[6]Mahuku G,Wangai A,Sadessa K,et al.. First report of Maize chlorotic mottle virus and maize lethal necrosis on maize in Ethiopia[J]. Plant Dis.,2015,99(12):1-4.
[7]Gell G,Sebestyén E,Balázs E. Recombination analysis of Sugercan mosaic virus(SCMV)in the Sugarcane mosaic virus(SCMV)subgroup of potyviruses[J]. Virus Genes,2015,50(1):79-86.
[8]王凤梅,李平路,孟庆华.玉米粗缩病抗病育种研究进展[J].种子,2017,36(2):45-49.Wang F M,Li P L,Meng Q H. Research progress on maize rough dwarf disease resistance breeding[J]. Seed,2017,36(2):45-49.
[9]李秀坤,刘昌林,周羽,等.玉米病毒病的研究进展[J].作物杂志,2015(3):13-16.Li X K,Liu C L,Zhou Y,et al.. Research progress of viral diseases in maize[J]. Crops,2015(3):13-16.
[10]张超,战斌慧,周雪平.我国玉米病毒病分布及危害[J].植物保护,2017,43(1):1-8.Zhang C,Zhan B H,Zhou X P. Distribution and damage of maize viral diseases in China[J]. Plant Protect.,2017,43(1):1-8.
[11]李帅,朱敏,夏子豪,等.云南省玉溪市玉米致死性坏死病毒原的分子鉴定[J].植物保护,2015,41(3):110-114Li S,Zhu M,Xia Z H,et al.. Molecular identification of the viruses causing maize lethal necrosis disease in Yunnan Province[J]. Plant Protect.,2015,41(3):110-114.
[12]王芳,高正良,周本国,等.利用小RNA高通量测序技术检测玉米病毒[J].植物病理学报,2017,47(3):422-427Wang F,Gao Z L,Zhou B G,et al.. Detection of viruses in maize by high-throughput sequencing of small RNA[J]. Acta Phytopathol. Sin.,2017,47(3):422-427.
[13]Zhang C L,Gao R,Wang J,et al.. Molecular variability of Tobacco vein banding mosaic virus populations[J]. Virus Res.,2011,158:188-198.
[14]Tamura K,Dudley J, Nei M, et al.. MEGA4:Molecular evolutionary genetics analysis(MEGA)software version 4. 0[J]. Mol. Biolo. Evol.,2007,24:1596-1599.
[15]翟玉山,赵贺,张海,等.甘蔗NAD(P)H脱氢酶复合体O亚基基因克隆及其与甘蔗花叶病毒VPg互作研究[J].作物学报,2019,2019:1-12.Zhai Y S,Zhao H,Zhang H,et al.. Cloning of NAD(P)H complex O subunit gene and its interaction with VPg of Sugarcane mosaic virus[J]. Acta Agron. Sin.,2019,2019:1-12.
[16]Adams M J,Antoniw J F,Beaudoin F. Overview and analysis of the polyprotein cleavage sites in the family Potyviridae[J].Mol. Plant Pathol.,2005,6(4):471-87.
[17]Cheng G,Dong M,Xu Q,et al.. Dissecting the molecular mechanism of the subcellular localization and cell-to-cell movement of the Sugarcane mosaic virus P3N-PIPO[J]. Sci.Rep.,2017,7(1):9868.
[18]Zhu M,Chen Y,Ding X S,et al.. Maize elongin C interacts with the viral genome-linked protein, VPg, of Sugarcane mosaic virus and facilitates virus infection[J]. New Phytol.,2014,203(4):1291-1304.
[19]Apriasti R, Widyaningrum S, Hidayati W N, et al.. Full sequence of the coat protein gene is required for the induction of pathogen-derived resistance against Sugarcane mosaic virus in transgenic sugarcane[J]. Mol. Biol. Rep.,2018,45(6):2749-2758.
[20]Tang W,Yan Z Y,Zhu T S,et al.. The complete genomic sequence of Sugarcane mosaic virus from Canna spp. in China[J]. Virol. J.,2018,15(1):147.
[21]吕庆雪,高嵩,何欢,等.新型玉米种子生产技术表达载体的构建及遗传转化[J].基因组学与应用生物学,2017(6):2497-2501.Lv Q X, Gao S, He H, et al.. Construction and genetic transformation of expression vector in new-type maize seeds production technology[J]. Genomics Appl. Biol.,2017(6):2497-2501.