以营养缺陷型筛选标记基因ura3为靶标,利用基于核糖核蛋白(ribonucleoprotein,RNP)的成簇规律间隔短回文重复序列及其相关蛋白9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9,CRISPR/Cas9)...以营养缺陷型筛选标记基因ura3为靶标,利用基于核糖核蛋白(ribonucleoprotein,RNP)的成簇规律间隔短回文重复序列及其相关蛋白9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9,CRISPR/Cas9)基因编辑技术,对灵芝(Ganoderma lucidum)双核菌株‘沪农灵芝1号’G0119及从其分离的2个交配亲和的单核菌株L1、L2进行基因编辑,比较单双核的编辑效率;对编辑的2个单核,进行杂交,以获得ura3被编辑的双核基因位点纯合菌株。结果表明:利用CRISPR/Cas9基因编辑技术,获得G0119双核编辑株7个,经双单杂交实验鉴定,有6个G0119编辑株为L1核被编辑,1个为L2核被编辑,7个编辑株均为杂合子;获得L1、L2的单核编辑株分别为10、7个,其中L1核被编辑的效率为100%。将突变类型均为C碱基缺失的单核L1-2和L2-1编辑株以及A插入碱基的L1-5和L2-3编辑株分别进行单单杂交,获得2株ura3被破坏的双核纯合菌株。利用CRISPR/Cas9基因编辑与杂交技术靶向创制灵芝基因位点纯合菌株的方法,可为创制灵芝其他功能基因的基因位点纯合菌株提供新的方法,也可为灵芝双核菌株的基因表型分析研究提供新的途径。展开更多
[Objective] This study was to design an intelligent greenhouse real-time monitoring system based on the core technology of Internet of Things in order to meet the needs of agricultural informatization and intellectual...[Objective] This study was to design an intelligent greenhouse real-time monitoring system based on the core technology of Internet of Things in order to meet the needs of agricultural informatization and intellectualization. [Method] Based on the application characteristics of Wireless Sensor Network （WSN）, the intelligent greenhouse monitoring system was designed. And for the incompleteness strategy of load balancing in the Low-Energy Adaptive Clustering Hierarchy （LEACH）, a Real- time Threshold Routing Algorithm （RTRA） was proposed. [Result] The performance of network lifetime and network delay of RTRA were tested in MATLAB and found that, within the same testing environment, RTRA can save nodes energy consumption, prolong network lifetime, and had better real-time performance than LEACH. The al- gorithm satisfies the crops＇ requirements on real-time and energy efficiency in the greenhouse system. [Conclusion] For the good performance on real-time, the de- signed intelligent greenhouse real-time monitoring system laid the foundation for the research and development of agricultural informatization and intellectualization.展开更多
文摘以营养缺陷型筛选标记基因ura3为靶标,利用基于核糖核蛋白(ribonucleoprotein,RNP)的成簇规律间隔短回文重复序列及其相关蛋白9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9,CRISPR/Cas9)基因编辑技术,对灵芝(Ganoderma lucidum)双核菌株‘沪农灵芝1号’G0119及从其分离的2个交配亲和的单核菌株L1、L2进行基因编辑,比较单双核的编辑效率;对编辑的2个单核,进行杂交,以获得ura3被编辑的双核基因位点纯合菌株。结果表明:利用CRISPR/Cas9基因编辑技术,获得G0119双核编辑株7个,经双单杂交实验鉴定,有6个G0119编辑株为L1核被编辑,1个为L2核被编辑,7个编辑株均为杂合子;获得L1、L2的单核编辑株分别为10、7个,其中L1核被编辑的效率为100%。将突变类型均为C碱基缺失的单核L1-2和L2-1编辑株以及A插入碱基的L1-5和L2-3编辑株分别进行单单杂交,获得2株ura3被破坏的双核纯合菌株。利用CRISPR/Cas9基因编辑与杂交技术靶向创制灵芝基因位点纯合菌株的方法,可为创制灵芝其他功能基因的基因位点纯合菌株提供新的方法,也可为灵芝双核菌株的基因表型分析研究提供新的途径。
基金Supported by the Science and Technology Surface Project of Yunnan Province(2010ZC142)the Doctoral Foundation of Dali University(KYBS201015),the Scientific Research Program for College Students of Dali University~~
文摘[Objective] This study was to design an intelligent greenhouse real-time monitoring system based on the core technology of Internet of Things in order to meet the needs of agricultural informatization and intellectualization. [Method] Based on the application characteristics of Wireless Sensor Network （WSN）, the intelligent greenhouse monitoring system was designed. And for the incompleteness strategy of load balancing in the Low-Energy Adaptive Clustering Hierarchy （LEACH）, a Real- time Threshold Routing Algorithm （RTRA） was proposed. [Result] The performance of network lifetime and network delay of RTRA were tested in MATLAB and found that, within the same testing environment, RTRA can save nodes energy consumption, prolong network lifetime, and had better real-time performance than LEACH. The al- gorithm satisfies the crops＇ requirements on real-time and energy efficiency in the greenhouse system. [Conclusion] For the good performance on real-time, the de- signed intelligent greenhouse real-time monitoring system laid the foundation for the research and development of agricultural informatization and intellectualization.
