生物技术通报2025，Vol.41Issue(3)：44-50,7.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0915
烟草染色体制备体系的建立与优化
Establishment and Optimization of a Tobacco Chromosome Preparation System
摘要
Abstract
[Objective]To develop a specialized chromosome preparation system for tobacco,laying the groundwork for advancements in tobacco chromosome engineering and the systematic exploration of distant hybridization research.[Method]The chromosome preparation system was established using the enzyme digestion method,with common tobacco as the test material.The experiment was designed to sample the root tip tissues at eight time intervals.Pre-treatment included four methods:A control group(without any pre-treatment),treatment with 20%wind medicated oil at room temperature for 2 h,exposure to an ice-water mixture at 0℃for 48 h,and N2O treatment,in which the N2O treatment was further divided into samples collected at 10-min intervals from 20 to 70 min;the root tip samples were enzymatically digest for different durations,including 35,45,and 55 min.Chromosome preparation was performed and observed for each treatment and duration,with the optimal method for tobacco chromosome preparation identified based on chromosome length,clarity,and dispersion.Merging improved processing methods and durations to establish a tobacco chromosome preparation system.The validation applicable to the method was further confirmed by preparing chromosome samples from various tobacco varieties.[Result]It is the optimal time between 10∶30 and 12∶30 for tobacco root tip sampling,30-40 min for N2O pretreatment and 45 min for enzymatic digestion were suitable for preparing chromosomes.High-quality chromosome samples were successfully obtained by applying this optimization system to three different germplasm resources:the ancestral species for common tobacco,common tobacco,and progeny derived from distant hybridization.[Conclusion]A preparation system for tobacco chromosomes is established,capable of preparing high-quality chromosome samples from tobacco root tips within 3 h.This system provides crucial technical support for research in distant hybridization and chromosome engineering.关键词
烟草/染色体制备/酶解法/种质资源/染色体工程Key words
tobacco/chromosome preparation/enzymatic hydrolysis/germplasm resources/chromosome engineering引用本文复制引用
韩梦荞,吴疆,李丽华,王昭懿,邓习,韦凤杰,任民,孙洋洋,李富欣..烟草染色体制备体系的建立与优化[J].生物技术通报,2025,41(3):44-50,7.基金项目
中国烟草总公司重大科技项目(110202201002(JY-02)),中国烟草总公司河南省公司创新项目(2024410000240025),中央级公益性科研院所基本科研业务费专项(1610232023022) （110202201002(JY-02）
