CRISPR/Cas技术在木本植物改良中的应用

侯静; 毛金燕; 翟惠; 王洁; 尹佟明

doi:10.12302/j.issn.1000-2006.202010017

侯静, 毛金燕, 翟惠, 王洁, 尹佟明

南京林业大学学报（自然科学版） ›› 2021, Vol. 45 ›› Issue (6) : 24-30.

PDF(1573 KB)

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

PDF(1573 KB)

南京林业大学学报（自然科学版） ›› 2021, Vol. 45 ›› Issue (6) : 24-30. DOI: 10.12302/j.issn.1000-2006.202010017

专题报道（执行主编施季森尹佟明陈金慧）

CRISPR/Cas技术在木本植物改良中的应用

作者信息 +

Application of CRISPR/Cas technique in woody plant improvement

Author information +

文章历史 +

摘要

木本植物育种周期长、种质资源匮乏,已成为限制木本植物种质创新的主要因素。CRISPR/Cas系统是近年来发展起来的能够实现对基因组进行精准定点编辑的技术,具有操作简单、周期短、效率高等优点,可实现基因的敲除、插入、替换等目的,从而精确地引入目标性状。目前,该技术已在多种家畜、昆虫、作物中得到了成功应用,对大量性状进行了改良,实现了种质资源的原始创新,大大缩短了育种年限。CRISPR/Cas技术的产生为木本植物的遗传改良提供了契机。笔者对CRISPR/Cas技术在杨树(Populus spp.)、苹果(Malus spp.)、柑橘(Citrus spp.)、葡萄(Vitis vinifera)、木薯(Cassava spp.)等木本植物育种中的应用进行了总结,该技术实现了T0代木本植物优良基因型的固定,在生长、材性、抗病性、抗旱性等性状上得到了明显改善,加快了木本植物育种进程,提高了育种效率。但该技术在木本植物中的应用尚处于起步阶段,还存在脱靶率高、编辑效率低、纯合突变少等问题。基于此,对CRISPR/Cas技术在木本植物中的应用前景进行了展望,以期为木本植物基因功能研究及品种改良提供有益参考。

Abstract

Long breeding periods and scarce germplasm resources have become the main bottlenecks in woody plant improvement. Recently, the CRISPR/Cas system has been developed into a precision site-directed editing technology that has been widely used in the breeding of woody plants such as Populus spp., Malus spp., Citrus spp., Vitis vinifera and Cassava spp.. The yield, quality, biotic- and abiotic-stress resistance, and other traits of woody plants have been significantly improved by genome editing, achieving the fixation of excellent genotypes within a single generation. This approach has accelerated the breeding process of woody plants and improves their breeding efficiency. Here, we summarize the application of CRISPR/Cas technology to woody plants, analyzing existing problems and future development trends. This review aims to provide a useful reference for gene function research and improvement of woody plants.

导出引用

侯静, 毛金燕, 翟惠, 等. CRISPR/Cas技术在木本植物改良中的应用[J]. 南京林业大学学报（自然科学版）. 2021, 45(6): 24-30 https://doi.org/10.12302/j.issn.1000-2006.202010017

HOU Jing, MAO Jinyan, ZHAI Hui, et al. Application of CRISPR/Cas technique in woody plant improvement[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2021, 45(6): 24-30 https://doi.org/10.12302/j.issn.1000-2006.202010017

中图分类号： Q943.2

参考文献

列表( 原文顺序 | 文献年度倒序 | 文中引用次数倒序 ) 可视化分析

[1]	万志兵, 戴晓港, 尹佟明林木遗传育种基础研究热点述评[J]. 林业科学, 2012, 48(2), 48:150-154. WAN Z B, DAI X G, YIN T M. Review on the hot topics of the basic studies for forest genetics and breeding[J]. Sci Silvae Sin, 2012, 48(2), 48:150-154. 本文引用 [1]

[2]	BEYING N, SCHMIDT C, PACHER M, et al. CRISPR-Cas9-mediated induction of heritable chromosomal translocations in Arabidopsis[J]. Nat Plants, 2020, 6(6):638-645.DOI: 10.1038/s41477-020-0663-x. https://doi.org/10.1038/s41477-020-0663-x https://doi.org/10.1038/s41477-020-0663-x 本文引用 [1]

[3]	SCHWARTZ C, LENDERTS B, FEIGENBUTZ L, et al. CRISPR-Cas9-mediated 75.5-Mb inversion in maize[J]. Nat Plants, 2020, 6(12):1427-1431.DOI: 10.1038/s41477-020-00817-6. https://doi.org/10.1038/s41477-020-00817-6 https://doi.org/10.1038/s41477-020-00817-6 本文引用 [1]

[4]	单奇伟, 高彩霞植物基因组编辑及衍生技术最新研究进展[J]. 遗传, 2015, 37(10), 37:953-973. SHAN Q W, GAO C X. Research progress of genome editing and derivative technologies in plants[J]. Hereditas, 2015, 37(10), 37:953-973.DOI: 10.16288/j.yczz.15-156. https://doi.org/10.16288/j.yczz.15-156 本文引用 [1]

[5]	PUCHTA H. Using CRISPR/Cas in three dimensions: towards synthetic plant genomes,transcriptomes and epigenomes[J]. Plant J, 2016, 87(1):5-15.DOI: 10.1111/tpj.13100. https://doi.org/10.1111/tpj.13100 http://doi.wiley.com/10.1111/tpj.2016.87.issue-1 本文引用 [1]

[6]	SHAN S, SOLTIS P S, SOLTIS D E, et al. Considerations in adapting CRISPR/Cas9 in nongenetic model plant systems[J]. Appl Plant Sci, 2020, 8(1):e11314.DOI: 10.1002/aps3.11314. https://doi.org/10.1002/aps3.11314 本文引用 [1]

[7]

CHEN

, WANG

, ZHANG

, et al. CRISPR/cas genome editing and precision plant breeding in agriculture[J]. Annu Rev Plant Biol, 2019, 70:667-697.DOI: 10.1146/annurev-arplant-050718-100049.

https://doi.org/10.1146/annurev-arplant-050718-100049

https://www.annualreviews.org/doi/10.1146/annurev-arplant-050718-100049

本文引用 [1]

[8]

VAN ZEIJL

, WARDHANI T A

, SEIFI KALHOR

, et al. CRISPR/Cas9-mediated mutagenesis of four putative symbiosis genes of the tropical tree Parasponia andersonii reveals novel phenotypes[J]. Front Plant Sci, 2018, 9:284.DOI: 10.3389/fpls.2018.00284.

https://doi.org/10.3389/fpls.2018.00284

http://journal.frontiersin.org/article/10.3389/fpls.2018.00284/full

本文引用 [1]

[9]	FAN D, LIU T, LI C, et al. Efficient CRISPR/Cas9-mediated targeted mutagenesis in Populus in the first generation[J]. Sci Rep, 2015, 5:12217.DOI: 10.1038/srep12217. https://doi.org/10.1038/srep12217 https://doi.org/10.1038/srep12217 本文引用 [1]

[10]	NISHITANI C, HIRAI N, KOMORI S, et al. Efficient genome editing in apple using a CRISPR/Cas9 system[J]. Sci Rep, 2016, 6:31481.DOI: 10.1038/srep31481. https://doi.org/10.1038/srep31481 https://doi.org/10.1038/srep31481 本文引用 [1]

[11]	ODIPIO J, ALICAI T, INGELBRECHT I, et al. Efficient CRISPR/Cas9 genome editing of phytoene desaturase in Cassava[J]. Front Plant Sci, 2017, 8:1780.DOI: 10.3389/fpls.2017.01780. https://doi.org/10.3389/fpls.2017.01780 http://journal.frontiersin.org/article/10.3389/fpls.2017.01780/full 本文引用 [1]

[12]	NAKAJIMA I, BAN Y, AZUMA A, et al. CRISPR/Cas9-mediated targeted mutagenesis in grape[J]. PLoS One, 2017, 12(5):e0177966.DOI: 10.1371/journal.pone.0177966. https://doi.org/10.1371/journal.pone.0177966 https://dx.plos.org/10.1371/journal.pone.0177966 本文引用 [1]

[13]	REN C, GUO Y, KONG J, et al. Knockout of VvCCD8 gene in grapevine affects shoot branching[J]. BMC Plant Biol, 2020, 20(1):41.DOI: 10.1186/s12870-020-2263-3. https://doi.org/10.1186/s12870-020-2263-3 https://doi.org/10.1186/s12870-020-2263-3 本文引用 [1]

[14]

FINLAYSON S

. Arabidopsis Teosinte Branched1-like 1 regulates axillary bud outgrowth and is homologous to monocot Teosinte Branched1[J]. Plant Cell Physiol, 2007, 48(5):667-677. DOI: 10.1093/pcp/pcm044.

https://doi.org/10.1093/pcp/pcm044

https://academic.oup.com/pcp/article-lookup/doi/10.1093/pcp/pcm044

本文引用 [1]

[15]

MUHR

, PAULAT

, AWWANAH

, et al. CRISPR/Cas9-mediated knockout of Populus BRANCHED1 and BRANCHED2 orthologs reveals a major function in bud outgrowth control[J]. Tree Physiol, 2018, 38(10):1588-1597.DOI: 10.1093/treephys/tpy088.

https://doi.org/10.1093/treephys/tpy088

https://academic.oup.com/treephys/article/38/10/1588/5110110

本文引用 [2]

[16]	田敏, 夏琼梅, 李纪元. 植物的次生生长及其分子调控[J]. 遗传, 2007, 29(11):1324-1330. TIAN M, XIA Q M, LI J Y. The secondary growth in plant and its molecular regulation[J]. Hereditas, 2007, 29(11):1324-1330.DOI: 10.16288/j.yczz.2007.11.006. https://doi.org/10.16288/j.yczz.2007.11.006 本文引用 [1]

[17]

TAKATA

, AWANO

, NAKATA M

, et al. Populus NST/SND orthologs are key regulators of secondary cell wall formation in wood fibers,phloem fibers and xylem ray parenchyma cells[J]. Tree Physiol, 2019, 39(4):514-525.DOI: 10.1093/treephys/tpz004.

https://doi.org/10.1093/treephys/tpz004

https://academic.oup.com/treephys/article/39/4/514/5365395

本文引用 [1]

[18]

YANG

, ZHAO

, RAN

, et al. PtoMYB156 is involved in negative regulation of phenylpropanoid metabolism and secondary cell wall biosynjournal during wood formation in poplar[J]. Sci Rep, 2017, 7:41209.DOI: 10.1038/srep41209.

https://doi.org/10.1038/srep41209

本文引用 [1]

[19]

ZHOU X

, JACOBS T

, XUE L

, et al. Exploiting SNPs for biallelic CRISPR mutations in the outcrossing woody perennial Populus reveals 4-coumarate:CoA ligase specificity and redundancy[J]. New Phytol, 2015, 208(2):298-301.DOI: 10.1111/nph.13470.

https://doi.org/10.1111/nph.13470

https://onlinelibrary.wiley.com/toc/14698137/208/2

本文引用 [1]

[20]	段硕. 柑橘易感溃疡病基因CsLOB1功能的研究[D]. 重庆:西南大学, 2017. DUAN S. Dissecting the function of susceptibility gene CsLOB1 of Citrus bacterial canker disease[D]. Chongqing:Southwest University, 2017. 本文引用 [1]

[21]

JIA

, ZHANG

, ORBOVI

$\acute{C}_{v}$

, et al. Genome editing of the disease susceptibility gene CsLOB1 in Citrus confers resistance to Citrus canker[J]. Plant Biotechnol J, 2017, 15(7):817-823.DOI: 10.1111/pbi.12677.

https://doi.org/10.1111/pbi.12677

https://onlinelibrary.wiley.com/doi/10.1111/pbi.12677

本文引用 [1]

[22]

PENG

, CHEN

, LEI

, et al. Engineering canker-resistant plants through CRISPR/Cas9-targeted editing of the susceptibility gene CsLOB₁ promoter in Citrus[J]. Plant Biotechnol J, 2017, 15(12):1509-1519.DOI: 10.1111/pbi.12733.

https://doi.org/10.1111/pbi.12733

http://doi.wiley.com/10.1111/pbi.2017.15.issue-12

本文引用 [1]

[23]	JIA H G, ORBOVIC V, WANG N. CRISPR-LbCas12a-mediated modification of Citrus[J]. Plant Biotechnol J, 2019, 17(10):1928-1937.DOI: 10.1111/pbi.13109. https://doi.org/10.1111/pbi.13109 https://onlinelibrary.wiley.com/toc/14677652/17/10 本文引用 [2]

[24]	郑小波. 疫霉菌及其研究技术[M]. 北京: 中国农业出版社, 1997. ZHENG X B. Phytophthora and its research technology[M]. Beijing: Chinese Agriculture Press, 1997. 本文引用 [1]

[25]	SHI Z, ZHANG Y, MAXIMOVA S N, et al. TcNPR3 from Theobroma cacao functions as a repressor of the pathogen defense response[J]. BMC Plant Biol, 2013, 13:204.DOI: 10.1186/1471-2229-13-204. https://doi.org/10.1186/1471-2229-13-204 https://doi.org/10.1186/1471-2229-13-204 本文引用 [1]

[26]

FISTER A

, LANDHERR

, MAXIMOVA S

, et al. Transient expression of CRISPR/Cas9 machinery targeting TcNPR3 enhances defense response in Theobroma cacao[J]. Front Plant Sci, 2018, 9:268.DOI: 10.3389/fpls.2018.00268.

https://doi.org/10.3389/fpls.2018.00268

http://journal.frontiersin.org/article/10.3389/fpls.2018.00268/full

本文引用 [1]

[27]

童蕴慧, 纪兆林, 徐敬友, 等. 灰霉病生物防治研究进展[J]. 中国生物防治, 2003, 19(3):131-135.

TONG Y

, JI Z

, XU J

, et al. Research progress on biological control of gray mold[J]. Chin J Biol Control, 2003, 19(3):131-135.DOI: 10.16409/j.cnki.2095-039x.2003.03.009.

https://doi.org/10.16409/j.cnki.2095-039x.2003.03.009

本文引用 [1]

[28]

黄幸, 丁峰, 彭宏祥, 等. 植物WRKY转录因子家族研究进展[J]. 生物技术通报, 2019, 35(12):129-143.

https://doi.org/10.13560/j.cnki.biotech.bull.1985.2019-0626

摘要

WRKY转录因子是植物中最大的转录调控因子家族之一,是调控植物许多生物过程信号网络的组成部分。WKRY转录因子具有多种生物学功能,在植物的生长发育和衰老、非生物和生物胁迫等过程中发挥着重要的作用。在DNA水平上,WRKY转录因子可与靶基因启动子中的W-box TTGAC（C/T）结合,通过自调节或交叉调节激活或抑制下游基因的表达调控其反应。在蛋白水平上,WRKY转录因子可以与多种蛋白相互作用,包括MAP激酶、组蛋白去乙酰化酶、抗性R蛋白、多种转录因子等,调节植物的生长发育或各种应激反应。对WRKY转录因子的结构特征、生物学功能、调控机制和网络等方面进行了综述,有助于更加全面了解其在植物中的作用。

HUANG

, DING

, PENG H

, et al. Research progress on family of plant WRKY transcription factors[J]. Biotechnol Bull, 2019, 35(12):129-143.DOI: 10.13560/j.cnki.biotech.bull.1985.2019-0626.

https://doi.org/10.13560/j.cnki.biotech.bull.1985.2019-0626

本文引用 [1] 摘要

WRKY transcription factors are one of the largest families of transcriptional regulators in plants and form an integral part of signaling webs that modulates many plant processes. WKRY transcription factors have a variety of biological functions and play an important role in plant growth,development and senescence,abiotic and biotic stress and so on. At DNA level,WRKY transcription factors can bind to W-box TTGAC（C/T）in the promoter of its target genes and activate or inhibit the expression of downstream genes to regulate their response by self-regulation or cross-regulation. At protein level,WRKY transcription factors can regulate plant growth and development or various stress responses by interacting with a variety of proteins,including MAP kinases,histone deacetylase,resistant R proteins,and a variety of transcription factors. This paper reviews the research progress on the structure,biological function,regulatory mechanism and network of WRKY transcription factors,which will help us to understand their roles in plants more comprehensively.

[29]	WANG X, TU M, WANG D, et al. CRISPR/Cas9-mediated efficient targeted mutagenesis in grape in the first generation[J]. Plant Biotechnol J, 2018, 16(4):844-855.DOI: 10.1111/pbi.12832. https://doi.org/10.1111/pbi.12832 http://doi.wiley.com/10.1111/pbi.2018.16.issue-4 本文引用 [2]

[30]	FEECHAN A, JERMAKOW A M, TORREGROSA L, et al. Identification of grapevine MLO gene candidates involved in susceptibility to powdery mildew[J]. Funct Plant Biol, 2008, 35(12):1255.DOI: 10.1071/fp08173. https://doi.org/10.1071/fp08173 http://www.publish.csiro.au/?paper=FP08173 本文引用 [1]

[31]	PESSINA S, LENZI L, PERAZZOLLI M, et al. Knockdown of MLO genes reduces susceptibility to powdery mildew in grapevine[J]. Hortic Res, 2016, 3:16016.DOI: 10.1038/hortres.2016.16. https://doi.org/10.1038/hortres.2016.16 https://doi.org/10.1038/hortres.2016.16 本文引用 [1]

[32]	MALNOY M, VIOLA R, JUNG M H, et al. DNA-free genetically edited grapevine and apple protoplast using CRISPR/Cas9 ribonucleoproteins[J]. Front Plant Sci, 2016, 7:1904.DOI: 10.3389/fpls.2016.01904. https://doi.org/10.3389/fpls.2016.01904 本文引用 [2]

[33]

GOMEZ M

, LIN Z

, MOLL

, et al. Simultaneous CRISPR/Cas9-mediated editing of cassava eIF4E isoforms nCBP-1 and nCBP-2 reduces cassava brown streak disease symptom severity and incidence[J]. Plant Biotechnol J, 2019, 17(2):421-434.DOI: 10.1111/pbi.12987.

https://doi.org/10.1111/pbi.12987

http://doi.wiley.com/10.1111/pbi.2019.17.issue-2

本文引用 [1]

[34]	鲁松. 干旱胁迫对植物生长及其生理的影响[J]. 江苏林业科技, 2012, 39(4):51-54. LU S. Effects of drought stress on plant growth and physiological traits[J]. J Jiangsu For Sci Technol, 2012, 39(4):51-54.DOI: 10.3969/j.issn.1001-7380.2012.04.015. https://doi.org/10.3969/j.issn.1001-7380.2012.04.015 本文引用 [1]

[35]	BRUNNER I, HERZOG C, DAWES M A, et al. How tree roots respond to drought[J]. Front Plant Sci, 2015, 6:547.DOI: 10.3389/fpls.2015.00547. https://doi.org/10.3389/fpls.2015.00547 本文引用 [1]

[36]

ZHOU

, ZHANG

, WANG

, et al. Root-specific NF-Y family transcription factor,PdNF-YB21,positively regulates root growth and drought resistance by abscisic acid-mediated indoylacetic acid transport in Populus[J]. New Phytol, 2020, 227(2):407-426.DOI: 10.1111/nph.16524.

https://doi.org/10.1111/nph.16524

https://onlinelibrary.wiley.com/toc/14698137/227/2

本文引用 [1]

[37]

, LIN Y

, WANG

, et al. The AREB1 transcription factor influences histone acetylation to regulate drought responses and tolerance in Populus trichocarpa[J]. Plant Cell, 2019, 31(3):663-686.DOI: 10.1105/tpc.18.00437.

https://doi.org/10.1105/tpc.18.00437

https://academic.oup.com/plcell/article/31/3/663-686/5985576

本文引用 [1]

[38]	JIAO Y N, WICKETT N J, AYYAMPALAYAM S, et al. Ancestral polyploidy in seed plants and angiosperms[J]. Nature, 2011, 473(7345):97-100.DOI: 10.1038/nature09916. https://doi.org/10.1038/nature09916 https://doi.org/10.1038/nature09916 本文引用 [1]

[39]	FRIEDLAND A E, TZUR Y B, ESVELT K M, et al. Heritable genome editing in C. elegans via a CRISPR-Cas9 system[J]. Nat Methods, 2013, 10(8):741-743.DOI: 10.1038/nmeth.2532. https://doi.org/10.1038/nmeth.2532 本文引用 [1]

[40]

ZETSCHE

, GOOTENBERG J

, ABUDAYYEH O

, et al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system[J]. Cell, 2015, 163(3):759-771.DOI: 10.1016/j.cell.2015.09.038.

https://doi.org/10.1016/j.cell.2015.09.038

https://linkinghub.elsevier.com/retrieve/pii/S0092867415012003

本文引用 [1]

[41]	COX D B T, GOOTENBERG J S, ABUDAYYEH O O, et al. RNA editing with CRISPR-Cas13[J]. Science, 2017, 358(6366):1019-1027.DOI: 10.1126/science.aaq0180. https://doi.org/10.1126/science.aaq0180 https://www.science.org/doi/10.1126/science.aaq0180 本文引用 [2]

[42]	HARRINGTON L B, BURSTEIN D, CHEN J S, et al. Programmed DNA destruction by miniature CRISPR-Cas14 enzymes[J]. Science, 2018, 362(6416):839-842.DOI: 10.1126/science.aav4294. https://doi.org/10.1126/science.aav4294 https://www.science.org/doi/10.1126/science.aav4294 本文引用 [2]

[43]	LU Y, YE X, GUO R, et al. Genome-wide targeted mutagenesis in rice using the CRISPR/Cas9 system[J]. Mol Plant, 2017, 10(9):1242-1245.DOI: 10.1016/j.molp.2017.06.007. https://doi.org/10.1016/j.molp.2017.06.007 https://linkinghub.elsevier.com/retrieve/pii/S1674205217301739 本文引用 [1]

[44]	MENG X, YU H, ZHANG Y, et al. Construction of a genome-wide mutant library in rice using CRISPR/Cas9[J]. Mol Plant, 2017, 10(9):1238-1241.DOI: 10.1016/j.molp.2017.06.006. https://doi.org/10.1016/j.molp.2017.06.006 https://linkinghub.elsevier.com/retrieve/pii/S1674205217301727 本文引用 [1]

[45]	ZHANG R, LIU J, CHAI Z, et al. Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing[J]. Nat Plants, 2019, 5(5):480-485.DOI: 10.1038/s41477-019-0405-0. https://doi.org/10.1038/s41477-019-0405-0 https://doi.org/10.1038/s41477-019-0405-0 本文引用 [1]

[46]	TIAN S, JIANG L, CUI X, et al. Engineering herbicide-resistant watermelon variety through CRISPR/Cas9-mediated base-editing[J]. Plant Cell Rep, 2018, 37(9):1353-1356.DOI: 10.1007/s00299-018-2299-0. https://doi.org/10.1007/s00299-018-2299-0 https://doi.org/10.1007/s00299-018-2299-0 本文引用 [1]

[47]

QIN

, LI J

, WANG Q

, et al. High-efficient and precise base editing of C·G to T·A in the allotetraploid cotton (Gossypium hirsutum) genome using a modified CRISPR/Cas9 system[J]. Plant Biotechnol J, 2020, 18(1):45-56.DOI: 10.1111/pbi.13168.

https://doi.org/10.1111/pbi.13168

https://onlinelibrary.wiley.com/toc/14677652/18/1

本文引用 [1]

[48]	LI C, ZHANG R, MENG X, et al. Targeted,random mutagenesis of plant genes with dual cytosine and adenine base editors[J]. Nat Biotechnol, 2020, 38(7):875-882.DOI: 10.1038/s41587-019-0393-7. https://doi.org/10.1038/s41587-019-0393-7 https://doi.org/10.1038/s41587-019-0393-7 本文引用 [1]

[49]	KUZMA J, GRIEGER K. Community-led governance for gene-edited crops[J]. Science, 2020, 370(6519):916-918.DOI: 10.1126/science.abd1512. https://doi.org/10.1126/science.abd1512 https://www.science.org/doi/10.1126/science.abd1512 本文引用 [1]

[50]	MA X, ZHANG X, LIU H, et al. Highly efficient DNA-free plant genome editing using virally delivered CRISPR-Cas9[J]. Nat Plants, 2020, 6(7):773-779.DOI: 10.1038/s41477-020-0704-5. https://doi.org/10.1038/s41477-020-0704-5 https://doi.org/10.1038/s41477-020-0704-5 本文引用 [1]

[51]

毛金燕, 翟惠, 王洁, 等. CRISPR/Cas技术及其作用机理[J/OL]. 分子植物育种:1-13.(2020 -11-18)[2021-02-27].

MAO

J Y

, ZHAI

, WANG

, et al. CRISPR/Cas technology and its action mechanisms[J/OL]. Molecular Plant Breeding:1-13 [2021-02-27]. http://kns.cnki.net/kcms/detail/46.1068.S.20201118.1049.004.html.

http://kns.cnki.net/kcms/detail/46.1068.S.20201118.1049.004.html.

本文引用 [1]