干旱区生态环境知识资源中心(Arid): Mesophyll protoplast isolation technique and flow cytometry analysis of ancient Platycladus orientalis (Cupressaceae)

Arid

DOI	10.3906/tar-1805-62
	Mesophyll protoplast isolation technique and flow cytometry analysis of ancient Platycladus orientalis (Cupressaceae)
	Zhou, Qianyi 1; Jiang, Zhaohong 2; Li, Yiming 1; Zhang, Tian 1; Zhu, Hailan 1; Zhao, Fei 3; Zhao, Zhong 1
通讯作者	Zhao, Zhong
来源期刊	TURKISH JOURNAL OF AGRICULTURE AND FORESTRY
ISSN	1300-011X
EISSN	1303-6173
出版年	2019
卷号	43 期号:3 页码:275-287
英文摘要	Platycladus orientalis (Cupressaceae) is a native tree species used widely for landscaping and afforestation in arid and semiarid areas in Asia. Ancient P. orientalis trees not only have an important humanistic and historical value but also have a significant scientific value in aging mechanism research. Mesophyll protoplasts are an important material for studies of plant regeneration, transient gene expression, and senescence. Although an easy and effective technique of mesophyll protoplast isolation from mature trees is urgently needed, the isolation parameters have great material specificity. The young tissue of plants is an ideal material for protoplast preparation. In this study, we employed an orthogonal experiment and several single-variable experiments to determine the main factors influencing the successful isolation of mesophyll protoplasts and established an efficient technique for isolating mesophyll protoplasts from the young scale leaves of ancient P. orientalis. The optimal parameters for mesophyll protoplast isolation are as follows: mechanical homogenization of the leaf tissue, 1.5% Cellulase R-10, 0.4% Macerozyme R-10, 0.4% Pectolyase Y-23, 1.0% ligninase, 0.7 M mannitol (pH 5.8), and 16 h of incubation. Two centrifugations (100 x g for 3 min and 500 x g for 5 min) were repeated 2 times to obtain purified protoplast suspension. The yield and viability of protoplasts under optimal conditions were 4.8 x 106 g FW-1 (per gram fresh weight) and 82.5%, respectively. The results of flow cytometry analysis showed that the isolated protoplasts had ideal viability to meet the demands of further protoplast-based research.
英文关键词	Flow cytometry mesophyll protoplast orthogonal experiment Platycladus orientalis protoplast isolation
类型	Article
语种	英语
国家	Peoples R China
开放获取类型	Bronze, Green Submitted
收录类别	SCI-E
WOS记录号	WOS:000471131900002
WOS关键词	SOMATIC EMBRYOGENESIS ; CALLUS FORMATION ; COTYLEDON PROTOPLASTS ; PINUS-PINASTER ; REGENERATION ; CULTURE ; LEAVES ; CELLS ; LEAF ; DIVISION
WOS类目	Agriculture, Multidisciplinary ; Agronomy ; Forestry
WOS研究方向	Agriculture ; Forestry
来源机构	西北农林科技大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/219136
作者单位	1.Northwest A&F Univ, Coll Forestry, Key Comprehens Lab Forestry, Yangling, Shaanxi, Peoples R China; 2.Northwest A&F Univ, Coll Life Sci, Yangling, Shaanxi, Peoples R China; 3.Beijing Agr Thchnol Extens Stn, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Zhou, Qianyi,Jiang, Zhaohong,Li, Yiming,et al. Mesophyll protoplast isolation technique and flow cytometry analysis of ancient Platycladus orientalis (Cupressaceae)[J]. 西北农林科技大学,2019,43(3):275-287.
APA	Zhou, Qianyi.,Jiang, Zhaohong.,Li, Yiming.,Zhang, Tian.,Zhu, Hailan.,...&Zhao, Zhong.(2019).Mesophyll protoplast isolation technique and flow cytometry analysis of ancient Platycladus orientalis (Cupressaceae).TURKISH JOURNAL OF AGRICULTURE AND FORESTRY,43(3),275-287.
MLA	Zhou, Qianyi,et al."Mesophyll protoplast isolation technique and flow cytometry analysis of ancient Platycladus orientalis (Cupressaceae)".TURKISH JOURNAL OF AGRICULTURE AND FORESTRY 43.3(2019):275-287.