CRISPR/Cas9系统在林木基因编辑中的应用

doi:10.16288/j.yczz.20-092

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Hereditas(Beijing) ›› 2020, Vol. 42 ›› Issue (7): 657-668.doi: 10.16288/j.yczz.20-092

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Application of CRISPR/Cas9 mediated gene editing in trees

Yingnan Chen^1,², Jing Lu^1,²

^1. Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing 210037, China
^2. Nanjing Forestry University, Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing 210037, China

Received:2020-04-04 Revised:2020-05-25 Online:2020-07-20 Published:2020-06-15
Supported by:
Supported by the Youth Elite Science Sponsorship Program by CAST No(YESS20160121);the Qing Lan Talent Support Program at Jiangsu Province, and the Priority Academic Program Development Program of Jiangsu Province

Abstract

Abstract:

The CRISPR/Cas9 system, which can induce precise modifications at a target gene, has been recognized as the most promising gene editing technology, and has played an important role in precision crop breeding. It also provides a new strategy for fundamental researches and molecular breeding of forest trees. Recently, CRISPR/Cas9-mediated gene editing has been applied more extensively in tree genetic studies. It has not only succeeded in developing new drought- resistance or disease-resistant cultivars, but also shows a great potential in regulating lignin biosynthesis and shortening the breeding cycle of forest trees. In this review, we summarize the application and advance of CRISPR/Cas9 in gene function identification and genetic improvement of forest plants. We also discuss the related problems and future perspectives. This review aims to provide a useful reference for tree functional genomics and genetic engineering breeding.

Key words: CRISPR/Cas9, gene editing, trees, gene function, genetic improvement

Yingnan Chen, Jing Lu. Application of CRISPR/Cas9 mediated gene editing in trees[J]. Hereditas(Beijing), 2020, 42(7): 657-668.

Figures/Tables 2

Table 1

Applications of CRISPR/Cas9 system in poplar"

研究目的	杨树种名	基因	sgRNA 数目	Cas 9密码子	参考文献
CRISPR 技术体系的建立与探索	毛白杨 (Populus tomentosa Carr. clone 741)	PtoPDS	4	human codon-optimized Cas9	[6]
	银灰杨 (P. tremula × alba clone 717-1B4) ; 欧美山杨 (P. tremula × tremuloides clone 353-38)	LEAFY、AGAMOUS	1~2	human codon-optimized Cas9	[8]
	银灰杨 (P. × canescens poplar clone INRA 717-1B4); 欧洲山杨 (P. tremula clone W52)	SOC1、SOC1 Paralog 1、SOC1 Paralog 2、AGL8.1、AGL8.2、NFP-like1-4、TOZ19	1~2	Cas9 wild type gene (Streptococcus pyogenes)	[9]
	新疆杨 [P. alba var. pyramidalis (P. bolleana)]	C2H2‐AZF	3	plant codon-optimized Cas9	[10]
木质素及类黄酮合成	银灰杨 (P. tremula × alba clone 717-1B4)	4CL1、4CL2	1	plant codon-optimized Cas9	[7]
	毛白杨 (P. tomentosa Carr. clone 741)	MYB115	3	plant codon-optimized Cas9	[14]
	毛白杨 (P. tomentosa Carr. clone 741)	PtoMYB156	3	plant codon-optimized Cas9	[15]
	毛白杨 (P. tomentosa Carr. clone 741)	PtoMYB170	3	plant codon-optimized Cas9	[16]
	毛白杨 (P. tomentosa Carr. clone 741)	PtrMYB57	3	plant codon-optimized Cas9	[17]
	毛白杨 (P. tomentosa Carr. clone 741)	JMJ25	1	plant codon-optimized Cas9	[18]
	毛白杨 (P. tomentosa Carr. clone 741)	MYB189	2	plant codon-optimized Cas9	[19]
木质部发育	毛白杨 (P. tomentosa Carr. clone 741)	PtoDWF4	3	plant codon-optimized Cas9	[20]
	欧美山杨 (P. tremula L.× P. tremuloides clone T89)	Pt×tvns09、vns10、 vns11、vns12	1	Arabidopsis codon-optimized Cas9	[12]
	毛白杨 (P. tomentosa Carr. clone 741)	PtoDET2	3	plant codon-optimized Cas9	[13]
生长与生长节律	银灰杨 (P. tremula × alba clone INRA 717-1B4)	PcBRC1、PcBRC2	2	plant codon-optimized Cas9	[11]
	银灰杨 (P. tremula × alba clone 717)	GNC	1	Arabidopsis codon-optimized Cas9	[21]
	银灰杨 (P. tremula × alba INRA clone 717-1B4)	LHY2	1	human codon-optimized Cas9	[22]
	欧美山杨 (P. tremula× tremuloides clone T89)	BRC1	2	maize-codon optimized Cas9	[23]
抗旱性状	毛果杨 (P. trichocarpa)	PtrADA2b-3	1	Arabidopsis codon-optimized Cas9	[24]
抗旱性状	84K杨 (P. alba × P. glandulosa)	PdNF-YB21	1	Arabidopsis codon-optimized Cas9	[25]
抗病性状	毛白杨 (P. tomentosa Carr. clone 741)	PtrWRKY18、PtrWRKY35	3	plant codon-optimized Cas9	[26]
酚甙类次生代谢产物	银灰杨 (P. tremula× alba INRA 717-1B4)	UGT71L1	1	Arabidopsis codon-optimized Cas9	[27]

Table 1

Table 2

Applications of CRISPR/Cas9 system in economic trees"

科	属	物种名称	外植体	遗传转化方法	靶基因	涉及性状	基因编辑效率(%)	参考文献
芸香科 (Rutaceae)	柑橘属 (Citrus)	甜橙 (Citrus sinensis cv. Valencia)	叶片	柑橘黄单胞菌 Xcc促进的农杆菌注射法	CsPDS	影响叶绿素生物合成,产生白化苗	3.2~3.9	[30]
		晚锦橙 (C. sinensis Osbeck)	上胚轴	农杆菌介导的遗传转化	CsLOB1 启动子	抗溃疡病	11.5~64.7	[31]
		葡萄柚 (C. paradisi)	上胚轴	农杆菌介导的遗传转化	CsLOB1 启动子	抗溃疡病	14.29~81.25	[33]
			上胚轴	农杆菌介导的遗传转化	CsLOB1	抗溃疡病	31.58~89.36	[34]
			上胚轴	农杆菌介导的遗传转化	CsLOB1	抗溃疡病	15~55	[36]
	枳属 (Poncirus)	枳橙 (Poncirus trifoliate L. Raf. × C. sinensis L. Osb.)	上胚轴	农杆菌介导的遗传转化	PDS	影响叶绿素生物合成,产生白化苗	45.5~75	[32]
	枳属 (Poncirus)	枳橙 (Poncirus trifoliate L. Raf. × C. sinensis L. Osb.)	上胚轴	农杆菌介导的遗传转化	Cs7g03360	叶片发育	15.55~79.67	[35]
猕猴桃科 (Actinidiaceae)	猕猴桃属 (Actinidia)	红阳猕猴桃 (Actinidia chinensis cv. Hongyang)	叶盘	农杆菌介导的遗传转化	AcPDS	影响叶绿素生物合成,产生白化苗	65.38~91.67	[38]
猕猴桃科 (Actinidiaceae)	猕猴桃属 (Actinidia)	中华猕猴桃‘Hort16A’ (A. chinensis cv. Hort16A)	叶盘	农杆菌介导的遗传转化	AcCEN4、 AcCEN	开花提前,果实早熟、紧凑	30~75	[40]
葡萄科 (Vitaceae)	葡萄属 (Vitis)	‘霞多丽’葡萄 (Vitis vinifera L. cv. Chardonnay)	悬浮细胞	农杆菌介导的遗传转化	IdnDH	降低酒石酸含量	100	[42]
		无核白葡萄 (V. vinifera cv. Thompson)	愈伤组织	农杆菌介导的遗传转化	VvWRKY52	抗灰霉病	31	[44]
		‘麝香’葡萄 (V. vinifera L. cv. Neo Muscat)	愈伤组织	农杆菌介导的遗传转化	VvPDS	产生白化植株	2.7~72.2	[43]
		‘葡萄砧木41B’ (V. vinifera cv. Chasselas × V. berlandieri)	胚性悬浮细胞	农杆菌介导的遗传转化	VvCCD8	增加分枝数	66.7	[45]
		‘霞多丽’葡萄 (V. vinifera L. cv. Chardonnay)	原生质体	PEG介导 RNP转化	MLO-7	抗白粉病	0.1	[46]
蔷薇科 (Rosaceae)	苹果属 (Malus)	‘金冠’苹果 (Malus prunifolia cv. Golden delicious)	原生质体	PEG介导 RNP转化	DIPM-1、DIPM-2、DIPM-4	抗白粉病	0.1~6.9	[46]
		‘苹果砧木JM2’ (M. prunifolia (Wild.) Borkh. ‘Seishi’?×?M. pumila Mill. var. paradisiaca Schneid. ‘M.9’)	叶盘	农杆菌介导的遗传转化	PDS	影响叶绿素生物合成,产生白化苗	31.8	[47]
		'嘎啦'苹果 (Malus × domestica Bork.)	嫩叶	农杆菌介导的遗传转化	MdPDS、MdTFL1.1	影响叶绿素生物合成,产生白化苗;也出现早花	85~93	[48]
	梨属 (Pyrus)	‘康弗伦斯’梨 (Pyrus communis L. cv. ‘Conference’)	嫩叶	农杆菌介导的遗传转化	PcTFL1.1	提前开花	9	[48]
石榴科 (Punicaceae)	石榴属 (Punica)	石榴 (Punica granatum L.)	毛状根	发根农杆菌遗传转化毛状根	PgUGT84A23、PgUGT84A24	降低酚类物质安石榴苷含量	-	[49]
茜草科 (Rubiaceae)	咖啡属 (Coffea)	中粒咖啡 (Coffea canephora clone 197)	愈伤组织	农杆菌介导的遗传转化	CcPDS	产生白化苗	30.4	[50]
梧桐科 (Sterculiaceae)	可可属 (Theobroma)	可可 (Theobroma cacao)	叶片	农杆菌介导的遗传转化	TcNPR3	抗疫霉菌	27	[51]
大戟科 (Euphorbiaceae)	木薯属 (Manihot)	木薯 (Manihot esculenta cv. 60444; cv. TME 204)	愈伤组织	农杆菌介导的遗传转化	MePDS	影响叶绿素生物合成,产生白化苗	97.1~98.9	[53]
		木薯 (M. esculenta cv. 60444)	愈伤组织	农杆菌介导的遗传转化	nCBP-1、 nCBP-2	抗木薯褐斑条纹病毒	91	[54]
		木薯 (M. esculenta)	愈伤组织	农杆菌介导的遗传转化	病毒基因AC2、AC3	抗非洲木薯花叶病毒	4.9~11	[55]
	橡胶树属 (Hevea)	橡胶树 (Hevea brasiliensis)	原生质体	PEG介导 RNP转化	FT、TFL1	提前开花	3.74~20.11	[57]
	麻疯树属 (Jatropha)	麻疯树 Jatropha curcas	子叶	农杆菌介导的遗传转化	JcCYP735A	生长和成花调控	-	[60]
大麻科 (Cannabaceae)	山豆麻属 (Parasponia)	Parasponia andersonii	茎和叶柄	农杆菌介导的遗传转化	PanHK4、PanEIN2、PanNSP1、PanNSP2	根瘤形成、茎形成层活性、植株性别	48~89	[61]
禾本科 (Poaceae)	簕竹属 (Bambusa)	绿竹 (Bambusa oldhamii)	原生质体	PEG介导质粒 DNA转化	PDS	影响叶绿素生物合成,产生白化苗	12.5	[62]
禾本科 (Poaceae)	牡竹属 (Dendrocalamus)	麻竹 (Dendrocalamus latiflorus Munro)	愈伤组织	农杆菌介导的遗传转化	DlmPSY1-A、DlmPSY1-B、DlmPSY1-C、GRG1	白化苗,株高增加	40~100	[63]

Table 2

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Application of CRISPR/Cas9 mediated gene editing in trees

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