陆均松(Dacrydium pierrei)染色体压片技术优化及核型分析  

Optimization and Karyotype Analysis of Dacrydium pierrei Chromosome Tableting Technology

作　　者：鹿文举 李兆基 宋希强[1] 霍少洁 赵莹[1] Lu Wenju;Li Zhaoji;Song Xiqiang;Huo Shaojie;Zhao Ying(Key Laboratory of Genetics and Germplasm Innovation of Tropical Characteristic Trees and Flowers,Key Laboratory of Biology of Tropical Characteristic Flowers and Trees,College of Forestry,Hainan University,Haikou,570228)

机构地区：[1]海南大学林学院,热带特色林木花卉遗传与种质创新教育部重点实验室,热带特色花木资源生物学重点实验室,海口570228

出　　处：《分子植物育种》2020年第15期5090-5096,共7页Molecular Plant Breeding

基　　金：海南省科协青年科技英才创新计划项目(QCXM201711)资助。

摘　　要：为了分析陆均松的核型特征,本研究在常规染色体压片方法的基础上,以海南陆均松(Dacrydium pierrei)茎尖为试材,从取材时间、预处理液的选择、处理时间、解离液的选择、解离时间和染色的时间上对染色体压片技术加以改进,然后利用Adobe Photoshop软件进行核型分析。结果表明,陆均松染色体压片最佳方法:取材时间为8:00 am^9:00 am,预处理使用0.1%秋水仙素处理2~4 h,固定液使用卡诺固定液(冰醋酸:无水酒精=1:3)在4℃下固定24 h,解离采用1 mol/L盐酸在水浴60℃下处理10 min,最后使用卡宝品红染液染色10 min。核型分析结果表明,陆均松染色体基数为x=10,染色体数目2n=20;其核型公式为2n=2x=20=14 m+6 sm,7对染色体为中部着丝点染色体类型(m),3对染色体为亚中着丝点染色体类型(sm)。本研究结果不仅为陆均松种内和种间杂交的研究提供理论依据,而且为陆均松染色体加倍等新品种的培育提供可靠的实验材料。This study was based on the conventional chromosome compression method,for analyzing the karyotype characteristics of Dacrydium pierrei,The stem tip of Dacrydium pierrei was used,which chromosome tableting technology was improved by picking time,selection of pretreatment liquid,treatment time,dissociation solution,dissociation time and dyeing time.Then we usd the Adobe Photoshop software for perform karyotyping.The results showed that the best method for the granulocyte compression of Dacrydium pierrei were as follows:the material was taken from 8:00 am to 9:00 am,the pretreatment was treated with 0.1% colchicine for 2 to 4 h,the fixative solution was carnot fixed solution(acetic acid:absolute alcohol=1:3) fixed at 4℃ for 24 h,and the dissociation was treated with 1 mol/L hydrochloric acid in water bath at 60℃for 10 min,finally dyed with carbo red dye solution for10 min.The karyotype analysis showed that the chromosome base of Dacrydium pierrei was x=10,and the number of chromosomes was 2 n=20;The karyotype formula was 2 n=2 x=20=14 m+6 sm,that seven pairs of chromosomes were the central centromere chromosome type(m),and three pairs of chromosomes were the sub-centralized chromosome type(sm).The results of this study provided not only the theoretical basis for the study of intraspecific and interspecific hybridization of Dacrydium pierrei,but also the reliable experimental materials for the breeding of new varieties such as doubled chromosomes of Dacrydium pierrei.

关 键 词：陆均松(Dacrydium pierrei) 染色体 压片技术 核型分析 

分 类 号：S791.24[农业科学—林木遗传育种]