女贞种质资源ISSR分析与抗寒优株选育  

ISSR cluster analysis and selection of superior cold resistant strain of Ligustrum lucidum germplasm resources

作　　者：姜楠南[1] 孙音[1] 刘凤栾[2] 王媛 卢洁 杨传祥 房义福[1] JIANG Nannan;SUN Yin;LIU Fengluan;WANG Yuan;LU Jie;YANG Chuanxiang;FANG Yifu(Landscape and Flower Institute,Shandong Academy of Forestry,Jinan 250014,Shandong,China;Shanghai Chenshan Botanical Garden,Shanghai Chenshan Plant Science Research Center of Chinese Sciences Academy,Shanghai 201602,China;Shandong Provincial Tree Seedling and Flower Station,Jinan 250014,Shandong,China;Taihe Garden Greening Co.Ltd.,Rizhao 276800,Shandong,China)

机构地区：[1]山东省林业科学研究院园林与花卉所,山东济南250014 [2]上海辰山植物园中国科学院上海辰山植物科学研究中心,上海201602 [3]山东省林木种苗和花卉站,山东济南250014 [4]泰禾园林绿化有限公司,山东日照276800

出　　处：《经济林研究》2021年第2期18-26,73,共10页Non-wood Forest Research

基　　金：山东省农业良种工程“常绿半常绿树种选育”(鲁科农社字2012lz12-03)。

摘　　要：【目的】筛选山东及其周边区域抗寒性状稳定的女贞优株。【方法】对来自不同地区的60份女贞种质进行连续7 a的大田抗寒表型观测,并对其进行分类。采用石蜡切片法,观察并比较初选越冬叶片保有率较高与叶片保有率较低的种质叶片解剖结构的差异。初冬,选取1年生枝为材料,在不同低温条件下处理后测定相对电导率,拟合Logistic方程,计算低温半致死温度。采用CTAB法,提取女贞幼叶基因组DNA,利用ISSR分子标记技术对女贞的基因组DNA进行多态性扩增,使用POPGENE软件计算引物的多态性指标,使用NTSYS软件进行种质间的UPGMA聚类分析。【结果】根据大田抗寒观测结果,将女贞抗寒表现由强到弱分为Ⅰ~Ⅴ级。越冬叶片保有率较高的女贞叶片的栅栏组织细胞排列更紧密,栅栏组织和海绵组织厚度的比值更高。女贞的低温半致死温度为-15.1~-4.2℃,其中A4的低温半致死温度最低。从96条引物中筛选出9条条带清晰多态性强的引物,共扩增出2013条带,多态性比率100%,Nei’s基因多样性指数为0.1163~0.1721;Shannon信息指数为0.2060~0.2953,在遗传相似系数0.298处60份女贞种质可分为抗寒性较强和较弱2类种质。【结论】优株A4的抗寒性强且稳定,是良好的抗寒女贞育种材料。供试女贞材料的ISSR聚类结果可反映其亲缘关系,也表明不能按照优株地理距离聚类。ISSR聚类结果与抗寒性表现相符,暗示亲缘关系近的女贞种质具有相似的抗寒表现。【Objective】Select Ligustrum lucidum superior plant with stable cold resistance traits in Shandong and its surrounding areas.【Method】The cold resistant phenotype of 60 materials from different regions were observed for 7 years and classified.The paraffin section method was used to observe and compare the differences in leaf anatomical structure of the germplasm with higher leaf retention rate and lower leaf retention rate after primary selection.In early winter,the annual branches were selected to determine the relative electrical conductivity by artificial cooling,the Logistic equation was used to calculate the half lethal temperature.Genomic DNA was extracted from young leaves following the CTAB method,and then were amplified by ISSR molecular marker technology.The percentage of polymorphic loci,the indexes of Nei’s genetic diversity and Shannon information were estimated by POPGENE.The cluster analysis of 60 germplasms of materials based on unweighted pair-group method and two populations based on Nei’s genetic distance were performed on NTSYS to classify germplasm materials.【Result】According to the field observation results of cold resistance,the cold resistance performance of L.lucidum was divided into grade I to grade V from strong to weak.Paraffin section showed that the palisade tissue cells arranged more closely and the ratio of palisade tissue thickness to sponge tissue thickness was higher.The semi-lethal temperature of L.lucidum was estimated to be between-15.1℃and-4.2℃by electrical conductivity,and the semi-lethal temperature of A4 was the lowest.Nine primers with clear and strong polymorphism bands were screened out from 96 primers,and a total of 2013 bands were amplified,the polymorphic ratio was 100%.The Nei’s genetic diversity indices and the Shannon information indices were observed at the range of 0.1163-0.1721 and 0.2060-0.2953 respectively.At the genetic similarity coefficient of 0.298,60 germplasms materials were divided into two types:strong cold resistance and weak col

关 键 词：女贞 抗寒性 叶片解剖结构 相对电导率 ISSR 

分 类 号：S687[农业科学—观赏园艺]