Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia

doi:10.7525/j.issn.1673-5102.2021.06.006

Abstract

Abstract:

In order to clarify the mechanism of MADS-box transcription factor gene family member in flowering and floral organ development in Macadamia integrifolia， aMiMADS-box gene was cloned from the leaves of M. integrifolia “GUIRE1” cultivar by PCR technique and analyzed its structure and function by Bioinformatics method. The cloned cDNA sequence of MiMADS-box gene was 1 179 bp in length， 729 bp open reading frame （ORF）， encoded 242 amino acids. MiMADS-box， an unstable hydrophilic protein that had no transmembrane structure， no signal peptide and localized in the nucleus， had MADS superfamily and K-box superfamily conserved domains belonging to the MIKCc-type family. Phylogenetic analysis clustered MiMADS-box and AGL7/AP1， AGL10/CAL1， AGL8/FUL， AGL79 into FUL-AP1（SQUA） subfamily. The transcriptome data were used to analyze the expression patterns of MiMADS-box gene in branch， flower and leave of M. integrifolia “GUIRE1” and “695” cultivar. Results indicated that MiMADS-box had the lowest expression in the flower of “GUIRE1”， but the expression level of “695” was the highest. The green fluorescent protein fusion vector pGREEN-MiMADS-box-ORF-GFP was constructed by homologous recombination. It was speculated that MiMADS-box was closely related to flowering and floral organ development. The study provided a theoretical and technical guide for clarification the role MiMADS-box in flowering regulation mechanism of M. integrifolia.

Key words: Macadamia integrifolia, MiMADS-box, gene cloning, vector construction

CLC Number:

S664.9

Qiu-Jin TAN, Tao ZHANG, Yuan-Rong WEI, Shu-Fang ZHENG, Xiu-Hua TANG, Wen-Lin WANG. Cloning and Expression Vector Construction of a Flowering Related Gene MiMADS-box from Macadamia integrifolia[J]. Bulletin of Botanical Research, 2021, 41(6): 888-895.

References

1	Borompichaichartkul C，Luengsode K，Chinprahast N，et al.Improving quality of macadamia nut（Macadamia integrifolia） through the use of hybrid drying process［J］.Journal of Food Engineering，2009，93（3）：348-353.
2	王文林，陈海生，张涛，等.澳洲坚果bHLH基因家族成员MibHLH48的结构与功能分析［J/OL］.分子植物育种，2020：1-16.（2020-08-05）..
2	Wang W L，Chen H S，Zhang T，et al.Structural and functional analysis of Macadamia integrifolia bHLH gene family member MibHLH48［J/OL］.Molecular Plant Breeding，2020：1-16.（2020-08-05）..
3	Ng M，Yanofsky M F.Function and evolution of the plant MADS-box gene family［J］.Nature Reviews Genetics，2001，2（3）：186-195.
4	Ratcliffe O J，Nadzan G C，Reuber T L，et al.Regulation of flowering in Arabidopsis by an FLC homologue［J］.Plant physiology，2001，126（1）：122-132.
5	Honma T，Goto K.Complexes of MADS-box proteins are sufficient to convert leaves into floral organs［J］.Nature，2001，409（6819）：525-529.
6	Pelaz S，Gustafson-Brown C，Kohalmi S E，et al.APETALA1 and SEPALLATA3 interact to promote flower development［J］.The Plant Journal，2001，26（4）：385-394.
7	Ma H，Yanofsky M F，Meyerowitz E M.AGL1-AGL6，an Arabidopsis gene family with similarity to floral homeotic and transcription factor genes［J］.Genes & Development，1991，5（3）：484-495.
8	Par?Enicova? L，De Folter S，Kieffer M，et al.molecular and phylogenetic analyses of the complete MADS-Box transcription factor family in Arabidopsis：new openings to the MADS World［J］.The Plant Cell，2003，15（7）：1538-1551.
9	Zhao Y F，Broholm S K，Wang F，et al.TCP and MADS-Box transcription factor networks regulate heteromorphic flower type identity in Gerbera hybrida［J］.Plant Physiology，2020，184（3）：1455-1468.
10	Montiel G，Gaudet M，Laurans F，et al.Overexpression of MADS-box gene AGAMOUS-LIKE 12 activates root development in Juglans sp.and Arabidopsis thaliana［J］.Plants，2020，9（4）：444.
11	Huang F Y，Zhang Y H，Hou X L.BcAP3，a MADS box gene，controls stamen development and male sterility in Pak-choi（Brassica rapa ssp. chinensis）［J］.Gene，2020，747：144698.
12	Yin X M，Liu X，Xu B X，et al.OsMADS18，a membrane-bound MADS-box transcription factor，modulates plant architecture and the abscisic acid response in rice［J］.Journal of Experimental Botany，2019，70（15）：3895-3909.
13	王文林，韦媛荣，郑树芳，等.澳洲坚果MYB1基因克隆及生物信息学分析［J］.南方农业学报，2020，51（2）：245-254.
13	Wang W L，Wei Y R，Zheng S F，et al.Cloning and bioinformatics analysis of MYB1 gene from Macadamia integrifolia［J］.Journal of Southern Agriculture，2020，51（2）：245-254.
14	樊松乐，王纪坤，谢贵水，等.R2R3型MYB转录因子HbMYB88结构和功能分析［J］.植物研究，2020，40（1）：106-116.
14	Fan S L，Wang J K，Xie G S，et al.Structural and functional analysis of R2R3 MYB transcription factor HbMYB88［J］.Bulletin of Botanical Research，2020，40（1）：106-116.
15	Wei M M，Wang Y J，Pan R R，et al.Genome-wide identification and characterization of MADS-box family genes related to floral organ development and stress resistance in Hevea brasiliensis Müll.Arg［J］.Forests，2018，9（6）：304.
16	Wang R Z，Ming M L，Li J M，et al.Genome-wide identification of the MADS-box transcription factor family in pear （Pyrus bretschneideri） reveals evolution and functional divergence［J］.PeerJ，2017，5：e3776.
17	Herbert S W，Walton D A，Wallace H M.The influence of pollen-parent and carbohydrate availability on macadamia yield and nut size［J］.Scientia Horticulturae，2019，251：241-246.
18	Herbert S W，Walton D A，Wallace H M.Pollen-parent affects fruit，nut and kernel development of Macadamia［J］.Scientia Horticulturae，2019，244：406-412.
19	Duduzile Buthelezi N M，Samukelo Magwaza L，Zeray Tesfay S.Postharvest pre-storage processing improves antioxidants，nutritional and sensory quality of Macadamia nuts［J］.Scientia Horticulturae，2019，251：197-208.
20	Becker A，Thei?en G.The major clades of MADS-box genes and their role in the development and evolution of flowering plants［J］.Molecular Phylogenetics and Evolution，2003，29（3）：464-489.
21	Arora R，Agarwal P，Ray S，et al.MADS-box gene family in rice：genome-wide identification，organization and expression profiling during reproductive development and stress［J］.BMC Genomics，2007，8：242.
22	Morel P，Chambrier P，Boltz V，et al.Divergent functional diversification patterns in the SEP/AGL6/AP1 MADS-Box transcription factor superclade［J］.The Plant Cell，2019，31（12）：3033-3056.
23	Chen L Y，Nan H Y，Kong L P，et al.Soybean AP1 homologs control flowering time and plant height［J］.Journal of Integrative Plant Biology，2020，62（12）：1868-1879.
24	Kong D Z，Shen X Y，Guo B，et al.Cloning and expression of an APETALA1-like gene from Nelumbo nucifera［J］.Genetics and Molecular Research，2015，14（2）：6819-6829.
25	Mandel M A，Yanofsky M F.The Arabidopsis AGL8 MADS box gene is expressed in inflorescence meristems and is negatively regulated by APETALA1［J］.The Plant Cell，1995，7（11）：1763-1771.
26	Huang F，Xu G L，Chi Y J，et al.A soybean MADS-box protein modulates floral organ numbers，petal identity and sterility［J］.BMC Plant Biology，2014，14：89.
27	Gao R M，Wang Y，Gruber M Y，et al.miR156/SPL10 modulates lateral root development，branching and leaf morphology in Arabidopsis by silencing AGAMOUS-LIKE 79［J］.Frontiers in Plant Science，2018，8：2226.
28	Zhou Y，Hu L F，Ye S F，et al.Overexpression of an APETALA1-like gene from cucumber（Cucumis sativus L.） induces earlier flowering and abnormal leaf development in transgenic Arabidopsis［J］.Canadian Journal of Plant Science，2019，99（2）：1-11.
29	黄海娇，李慧玉，姜静.BpAP1转基因白桦中开花相关基因的时序表达［J］.东北林业大学学报，2017，45（1）：1-6.
29	Huang H J，Li H Y，Jiang I.Quantitative expression analysis of several flowering-related genes in BpAP1 transgenic birch（Betula platyphylla × Betula pendula）［J］.Journal of Northeast Forestry University，2017，45（1）：1-6.
30	李爽，郑唐春，臧丽娜，等.小黑杨PsnAP1基因转化烟草的研究［J］.安徽农业科学，2014，42（8）：2278-2281，2308.
30	Li S，Zheng T C，Zang L N，et al.Transformation of tobaccos with Two AP1 genes isolated from poplar（Populus simonii × Populus nigra）［J］.Journal of Anhui Agricultural Sciences，2014，42（8）：2278-2281，2308.
31	王企，陈利娜，夏小丛，等.核桃FRUITFULL（JrFUL）基因的克隆及表达分析［J］.果树学报，2019，36（8）：992-1004.
31	Wang Q，Chen L N，Xia X C，et al.Cloning and expression analysis of walnut FRUITFULL（JrFUL） gene［J］.Journal of Fruit Science，2019，36（8）：992-1004.

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