流式细胞仪检测香花油茶、越南油茶基因组大小方法的建立及应用  

Establishment of a Flow Cytometry Method for Genome Size Deter-mination of Camellia osmantha and Camellia vietnamensis and Its Ap-plication

作　　者：吴方圆 蔡娅 郝丙青 贾艳霞[2] 叶航[1] 张照远[1] 马锦林[1] WU Fangyuan;CAI Ya;HAO Bingqing;JIA Yanxia;YE Hang;ZHANG Zhaoyuan;MA Jinlin(Guangxi Forestry Research Institute/Guangxi Key Laboratory of Special Non-wood Forest Cultivation and Utilization/Improved Variety and Cultivation Engineering Research Center of Oil-tea Camellia in Guangxi,Nanning,Guangxi 530002,China;Kunming Institute of Botany,Chinese Academy of Sciences/Molecular Biology Experiment Center,Germplasm Bank of Wild Species in Southwest China,Kunming,Yunnan 650000,China)

机构地区：[1]广西壮族自治区林业科学研究院/广西经济林培育与利用重点实验室/广西油茶良种与栽培技术创新中心,广西南宁530002 [2]中国科学院昆明植物研究所/中国西南生物种质资源库分子生物学实验中心,云南昆明650000

出　　处：《热带作物学报》2023年第8期1542-1550,共9页Chinese Journal of Tropical Crops

基　　金：广西林科院基本科研业务费专项(林科202109号);广西特色经济林培育与利用重点实验室自主课题资助项目(No.19-A-01-01);广西科学研究与技术开发计划项目(桂科AB21076012)。

摘　　要：为香花油茶(Camellia osmantha)、越南油茶(C. vietnamensis)优良品系基因组大小遗传多样性分析及特异种质筛选、倍性差异分析提供参考,针对解离液种类、内参植物、试验材料组织器官等关键环节,以细胞核解离情况、次生代谢物质降解情况、细胞峰出峰稳定情况、主峰清晰程度、主杂峰分离程度、取材容易程度、内参与待测样品基因组大小差异情况、基因组大小测定准确性及变异系数等方面为评价指标,构建基于香花油茶良种义禄(Camellia osmantha var. Yilu)、越南油茶优良品系鸿鸪23(C. vietnamensis var. Hongu 23)为试材的流式细胞术基因组大小检测技术体系,并用于香花油茶、越南油茶优良品系的测定。结果表明:研究发现4种常见的解离液WPB、Tris-MgCl2、OTTO、mGb中WPB、mGb解离效果较好,内参植物以豌豆、玉米为优先,试验材料组织器官以嫩叶或根尖为宜;香花油茶优良品系基因组大小在5.12~9.31 Gb之间变化,均值为7.60 Gb,1号、37号、45号种质在基因组大小分布上存在极端性,推测其存在倍性差异或其亲本间存在种间杂交;越南油茶优良品系基因组大小在9.17~9.85Gb之间变化,均值为9.47 Gb。香花油茶与越南油茶优良品系群体间在基因组大小上存在明显差异。A technique system for detecting the genome size of Camellia oleifera by flow cytometry was established to analyze the genome size genetic diversity between the superior lines of C.osmantha var.Yilu and C.vietnamensis var.Hongu 23,and to provide reference for selection of special germplasm and analysis of ploidy difference,based on the key links such as the type of dissociated liquid,the plant of internal reference,the tissue organ of the test material,evaluation indexes such as the nuclear dissociation,the degradation of secondary metabolites,the stability of the cell peak,the clear degree of the main peak,the separation degree of the main and miscellaneous peak,the easy degree of sampling, the difference of the genome size of the internal participating samples, the accuracy of genome size determi-nation and the coefficient of variation. The method was used to determine the genome size of the superior lines of C. osmantha and C. vietnamensis. The disintegrating effect of WPB and mGb was better among common dissociative fluids WPB, Tris-MgCl2, OTTO, mGb. Pean and corn were the favorable internal plants, and young leaf or root tip were the favorable experimental materials. The genome size of the superior lines of C. osmantha was from 5.12 to 9.31 Gb, with an average value of 7.60 Gb, the genome size distribution of the superior lines No. 1, No. 37 and No. 45 was extreme, and it was speculated that there were ploidy differences or interspecific hybridization between the parents. The genome size of superior lines of C. vietnamensis varied from 9.17 to 9.85 Gb, the mean value was 9.47 Gb. There was signifi-cant difference in genome size between the superior lines of C. osmantha and C. vietnamensis.

关 键 词：油茶 基因组大小 流式细胞仪 

分 类 号：S794.4[农业科学—林木遗传育种]