摘要
采用PCR和巢式PCR技术对6个新引进果蔗品种黔蔗08-1497,黔糖5号,云南红皮,广东黄皮,黔蔗08-688,黔糖3号和广东主栽果蔗品种黑皮果蔗(Badila)进行宿根矮化病菌检测,并对其中广东黄皮、云南红皮、黔糖3号及黑皮果蔗(Badila)的巢式PCR第二轮扩增产物(编号依次为Lxx-Gd-gz1、Lxx-Gd-gz2、Lxx-Gd-gz3及Lxx-Gd-gz4)进行核苷酸序列测定及分析。结果表明,PCR检测,仅黔蔗08-688甘蔗宿根矮化病菌检出率为20%(1/5),其余6个果蔗品种的宿根矮化病菌检出率均为0%;巢式PCR检测,黔蔗08-1497及黔糖5号宿根矮化病菌检出率为0%,广东黄皮检出率为75%;云南红皮、黔蔗08-688、黔糖3号及黑皮果蔗(Badila)检出率均为100%。Lxx-Gd-gz1、Lxx-Gd-gz2、Lxx-Gd-gz3及Lxx-Gd-gz4基因组相应区段核苷酸序列同源率为100%,均与巴西、澳大利亚、美国路易斯安娜州、中国云南、中国福建及广东湛江株系核苷酸序列同源率均为99%。表明该病菌遗传稳定性极高,变异极小。
PCR and nested-PCR were used to detect Leifsonia xyli subsp. xyli(Lxx),the causal bacterium of sugarcane stunting disease(RSD) were collected from seven different cultivars of fruit cane,including Qianzhe 08-1497,Qiantang No. 5,Yunnan hongpi,Guangdong huangpi,Qianzhe 08-688,Qiantang No. 3 and Badila,and the nucleotide sequence determination and analysis in fruit cane cultivars of Guangdong huangpi,Yunnan hongpi,Qiantang No. 3 and Badila(isolate number followed Lxx-Gd-gz1,Lxx-Gd-gz2,Lxx-Gd-gz3 and LxxGd-gz4) amplified products of nested-PCR in the second round were made. The results showed that RSD bacteria detection rate was 20 %(1 /5) only in Qianzhe 08-688,and Lxx detection rate was 0 % in the remaining six fruit cane cultivars by using PCR detection; However,Lxx detection rate was 0 % only in Qianzhe 08-1497 and Qiantang No. 5,Guangdong huangpi detection rate was 75 %; Yunnan hongpi,Qianzhe 08-688,Qiantang No. 3 and Badila detection rate was 100 % by nested-PCR detection. The intergenic transcribed spacer(ITS) region of 16s-23 s r DNA of Lxx-Gd-gz1,Lxx-Gd-gz2,Lxx-Gd-gz3 and Lxx-Gd-gz4 isolates shared almost 100 % nucleotide identity each other and shared almost 99 % nucleotide identity with Brazil,Australia,USA(Luisanna),China(Yunnan Province,Fujian Province and Zhanjiang City,Guangdong Province) isolates. It was that the genetic stability of Lxx was extremely high,and the variation was minimal.
出处
《西南农业学报》
CSCD
北大核心
2016年第5期1046-1051,共6页
Southwest China Journal of Agricultural Sciences
基金
广东省公益研究与能力创新专项资金项目(2014A020208094)
华南农业大学人才引进启动资金项目(4100k13009)
关键词
果蔗
宿根矮化病菌
PCR
巢式PCR
序列分析
Sugarcane
Leifsonia xyli subsp.xyli
PCR detection
Nested-PCR detection
Nucleotide sequence analysis