Using genetic and crispr/cas9 methods to achieve quadruple mutant in trichome trimeric complex for future study in fiber initiation mechanism | [Pakistan Journal of Botany • 2024]

Author(s):

1. TRINH NGOC AI: Tra Vinh University,No. 126 Nguyen Thien Thanh Street, Ward 5, Tra Vinh City,Viet Nam

2. NGHI KHAC NHU: Tra Vinh University,No. 126 Nguyen Thien Thanh Street, Ward 5, Tra Vinh City,Viet Nam

3. ANH PHU NAM BUI: Institute of Applied Technology, Thu Dau Mot University,Binh Duong Province,Vietnam

Abstract:

Despite many similarities between the molecular machinery for initiation in cotton fiber and Arabidopsis trichome, there are significant distinctions in how fiber and trichome pattern. While trichome initiation are clearly specified, there is a randomness in fiber initiation in seed coat. While characterizing the cotton fiber genes, the corresponding Arabidopsis gene is replaced by the cotton homolog, and all the complemented lines showed a defined trichome pattern similar to wild type Arabidopsis. Owing to the lack of appropriate tools, the fundamental dissimilarities in the patterning process remain unresolved. To completely understand the cotton fiber initiation mechanism and the difference in pattern formation between cotton fiber and Arabidopsis trichome, we aimed to create a cotton trimeric complex MYB2-DEL65/DEL61-TTG3 in Arabidopsis to study the fiber initiation mechanism. To achieve this ultimate objective, the first step is to generate Arabidopsis quadruple mutant (gl1gl3egl3ttg1). In this paper we successfully applied genetic and CRISPR/Cas9 approaches to obtain the quadruple mutant. Further, we also introduced the cotton MYB2-A gene into the quadruple mutant. The material generated by this paper will subsequently be used to decipher the molecular basis for cotton fiber initiation.

Page(s): 517-522

DOI: 10.30848/PJB2024-2(6)

Published: Journal: Pakistan Journal of Botany, Volume: 56, Issue: 2, Year: 2024

Keywords:

CRISPRCas9 , Trimeric complex , gl1gl3egl3ttg1 , Fiber initiation , MYB2DEL65 , Quadruple mutant , DEL61TTG3

References:

[1] Bortesi L.,Fischer R. .2015 .The CRISPR/Cas9 system for plant genome editing and beyond. Biotechnol, 33(1) : 41-52.

[2] Clough S.J.,Bent A.F. .1998 .Floral dip: a simplified method forAgrobacterium-mediated transformation of Arabidopsis thaliana. The Plant, 00343(6) : 1365-313x.

[3] Cox N.,Smith L.M. .2019 .A novel upstream regulator of trichome development inhibitors. Plant, 10(4) : 01269-1400.

[4] Doroshkov A.V.,Konstantinov D.K.,Afonnikov D.A.,Gunbin K.V. .2019 .The evolution of gene regulatory networks controlling Arabidopsis thaliana L. trichome development. BMC Plant Biol, 10(1) : 53.

[5] Esch J.J.,Chen M.,Sanders M.,Hillestad M.,Ndkium S.,Idelkope B.,Marks M.D. .2003 .A contradictory GLABRA3 allele helps define: Interactions controlling trichome development in Arabidopsis. Development, 10(24) : 5885-5894.

[6] Fambrini M.,Pugliesi C. .2019 .The Dynamic GeneticHormonal Regulatory Network Controlling the Trichome Development in Leaves. , 8(8) : 253.

[7] Galway M.E.,Masucci J.D.,Lloyd A.M.,Walbot V.,Davis R.W.,Schiefelbein J.W. .1994 .The TTG gene is required to specify epidermal cell fate and cell patterning in the Arabidopsis root. Develop. Biol, 166(2) : 740-754.

[8] Glover B.J. .2000 .Differentiation in plant epidermal cells. J. Exp. Bot., 10(344) : 497-505.

[9] Guan X.,Pang M.,Nah G.,Shi X.,Ye W.,Stelly D.M.,Chen Z.J. .2014 .miR828 and miR858 regulate homoeologous MYB2 gene functions in Arabidopsis trichome and cotton fibre development. , 10 : .

[10] Hahn F.,Mantegazza O.,Greiner A.,Hegemann P.,Eisenhut M.,Weber A.P.M. .2017 .An efficient visual screen for CRISPR/Cas9 activity in Arabidopsis thaliana. Front. Plant Sci., 10 : 39.

[11] Hulskamp M. .2004 .Plant trichomes: a model for cell differentiation. Nat. Rev. Mol. Cell Biol, 5(6) : S1-480.

[12] Humphries J.,Walker A.,Timmis J.,Orford S. .2005 .Two WD-repeat genes from cotton are functional homologues of the Arabidopsis thaliana TRANSPARENT TESTA GLABRA1 (TTG1) gene. Plant Mol. Biol, 10(1) : 67-81.

[13] Hussain B.,Lucas S.J.,Budak H. .2018 .CRISPR/Cas9 in plants: At play in the genome and at work for crop improvement. Briefings Fun, 10(5) : 319-328.

[14] Junker A.,M.M. Muraya K.,Weigelt-Fischer F.,AranaCeballos C.,Klukas A.E.,Melchinger T.,Altmann T. .2014 .Optimizing experimental procedures for quantitative evaluation of crop plant performance in high throughput phenotyping systems. , 5 : .

[15] Koornneef M. .1981 .The complex syndrome of ttg mutants. Arab, 18 : 45-51.

[16] Larkin J.C.,M.L. Brown J.,Schiefelbein J. .2003 .How do cells know what they want to be when they grow up? Lessons from epidermal patterning in Arabidopsis. , 54(1) : 403-430.

[17] Li F.,Fan G.,Lu C.,Zou C.,Kohel R.J.,Yu S. .2015 .Genome sequence of cultivated upland cotton (Gossypium hirsutum TM-1) provides insights into genome evolution. Nat, 33(5) : 524-530.

[18] Li F.,Fan G.,Wang K.,Sun F.,Yuan Y.,Song G.,Yu S. .2014 .Genome sequence of the cultivated cotton Gossypium arboreum. Nat. Genet, 10(6) : 567-572.

[19] Liu J.,Jiao T. J.,Lu F.,Xu F. .2017 .Trichomes as a natural biophysical barrier for plants and their bioinspired applications. Soft Matter, 13(30) : 5096-5106.

[20] Morohashi K.,Zhao M.,Yang M.,Read B.,Lloyd A.,Lamb R.,Grotewold E. .2007 .Participation of the Arabidopsis bHLH factor gl3 in trichome initiation regulatory events. Plant, 145(3) : 736-746.

[21] Oppenheimer D.G.,Herman P.L.,Sivakumaran S.,Esch J.,Marks M.D. .1991 .A myb gene required for leaf trichome differentiation in Arabidopsis is expressed in stipules. Cell, 67(3) : 483-493.

[22] Payne C.T.,Zhang F.,Lloyd A.M. .2000 .GL3 encodes a bHLH protein that regulates trichome development in Arabidopsis through interaction with GL1 and TTG1. , 156(3) : 1349-1362.

[23] Pesch M.,Hülskamp M. .2009 .One, two, three models for trichome patterning in Arabidopsis? Curr. Opin. , 12(5) : 587-592.

[24] Ramsay N.A.,Glover B.J. .2005 .MYB-bHLH-WD40 protein complex and the evolution of cellular diversity. Trends in Plant Sci., 10(2) : 63-70.

[25] Sato Y.,Shimizu-Inatsugi R.,Yamazaki M.,Shimizu K.K.,Nagano A.J. .2019 .Plant trichomes and a single gene GLABRA1 contribute to insect community composition on field-grown Arabidopsis thaliana. BMC Plant Biol, 10(1) : 163.

[26] Schiefelbein J. .2003 .Cell-fate specification in the epidermis: a common patterning mechanism in the root and shoot. Curr. Opin. Plant Biol, 6(1) : 74-78.

[27] Serna L. .2004 .A network of interacting factors triggering different cell fates. The Plant Cell, 10(9) : 2258-2263.

[28] Serna C.,Martin C. .2006 .Trichomes: different regulatory networks lead to convergent structures. Trends Plant Sci., 11(6) : 274-280.

[29] Tang T.,Yu X.,Yang H.,Gao Q.,Ji H.,Wang Y.,Dai C. .2018 .Development and validation of an effective CRISPR/Cas9 vector for efficiently isolating positive transformants and transgene-free mutants in a wide range of plant species. Front. Plant Sci., 01533 : 1533-1533.

[30] Walker A.R.,Davison P.A.,Bolognesi-Winfield A.C.,James C.M.,Srinivasan N.,Blundell T.L.,Gray J.C. .1999 .The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein. The Plant Cell, 11(7) : 1337-1350.

[31] Wang G.,Zhao G.H.,Jia Y.H.,Du X.M.,Integ X.M. .2013 .Identification and Characterization of Cotton Genes Involved in FuzzFiber Development. , 10(7) : 619-630.

[32] Wang K.,Wang Z.,Li F.,Ye W.,Wang J.,Song G.,Yu S. .2012 .The draft genome of a diploid cotton Gossypium raimondii. Nat Genet, 44(10) : 1098-1103.

[33] Wang S.,Wang J.W.,Yu N.,Li C.H.,Luo B.,Gou J.Y.,Chen X.Y. .2004 .Control of plant trichome development by a cotton fiber MYB gene. The Plant Cell, 16(9) : 2323-2334.

[34] Wang Z.,Yang Z.,Li F. .2019 .Updates on molecular mechanisms in the development of branched trichome in Arabidopsis and nonbranched in cotton. Plant, 10(9) : 1706-1722.

[35] Zhang F.,Gonzalez A.,Zhao M.,Payne C.T.,Lloyd A. .2003 .A network of redundant bHLH proteins functions in all TTG1-dependent pathways of Arabidopsis. Development, 10(20) : 4859-4869.

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