Construction of Suppression Expression Vector and Genetic Transformation of BpSPL9 gene from Betula platyphylla

doi:10.7525/j.issn.1673-5102.2023.01.004

Abstract

Abstract:

To clarify the function of SPL gene in growth and development of Betula platyphylla，the BpSPL9 gene was cloned， and the expression vector of BpSPL9 was constructed by CREST technology， and the genetic transformation was carried out by Agrobacterium mediated method， and the phenotype of the transgenic lines was observed， respectively. The results showed that the transgenic lines with BpSPL9 suppressed were successfully obtained. The height of transgenic lines was significantly lower than that of non-transgenic lines， with shorter node spacing and smaller leaf area. The contents of 6-BA and IAA in transgenic lines were lower than those of the non-transgenic lines. It is speculated that BpSPL9 gene participated in plant growth and development by affecting the synthesis of auxin and cell division.

Key words: Betula platyphylla, SPL9, CREST, genetic transformation

CLC Number:

S792.153

Jinxia DU, Tingting SHEN, Haoran WANG, Yiping LIN, Huiyu LI, Lianfei ZHANG. Construction of Suppression Expression Vector and Genetic Transformation of BpSPL9 gene from Betula platyphylla[J]. Bulletin of Botanical Research, 2023, 43(1): 30-35.

References

1	OHTA M， MATSUI K， HIRATSU K，et al.Repression domains of class II ERF transcriptional repressors share an essential motif for active repression［J］.The Plant Cell，2001，13（8）：1959-1968.
2	YANG Z， TIAN L N， LATOSZEK-GREEN M，et al. Arabidopsis ERF4 is a transcriptional repressor capable of modulating ethylene and abscisic acid responses［J］.Plant Molecular Biology，2005，58（4）：585-596.
3	YANG J H， ZHANG M F， YU J Q.Relationship between cytoplasmic male sterility and SPL-like gene expression in stem mustard［J］.Physiologia Plantarum，2008，133（2）：426-434.
4	YANG Z F， WANG X F， GU S L，et al.Comparative study of SBP-box gene family in Arabidopsis and rice［J］.Gene，2008，407（1-2）：1-11.
5	NING K， CHEN S， HUANG H J，et al.Molecular characterization and expression analysis of the SPL gene family with BpSPL9 transgenic lines found to confer tolerance to abiotic stress in Betula platyphylla Suk.［J］.Plant Cell，Tissue and Organ Culture（PCTOC），2017，130（3）：469-481.
6	LI L， SHI F， WANG Y Q，et al. TaSPL13 regulates inflorescence architecture and development in transgenic wheat（Triticum aestivum L.）［J］.Plant Science，2020，296：110516.
7	LIANG X W， NAZARENUS T J， STONE J M.Identification of a consensus DNA-binding site for the Arabidopsis thaliana SBP domain transcription factor，AtSPL14，and binding kinetics by surface plasmon resonance［J］.Biochemistry，2008，47（12）：3645-3653.
8	UNTE U S， SORENSEN A M， PESARESI P，et al. SPL8，an SBP-box gene that affects pollen sac development in Arabidopsis ［J］.The Plant Cell，2003，15（4）：1009-1019.
9	GANDIKOTA M， BIRKENBIHL R P， H?HMANN S，et al.The miRNA156/157 recognition element in the 3' UTR of the Arabidopsis SBP box gene SPL3 prevents early flowering by translational inhibition in seedlings［J］.The Plant Journal，2007，49（4）：683-693.
10	GOU J Y， FELIPPES F F， Liu C J，et al.Negative regulation of anthocyanin biosynthesis in Arabidopsis by a miR156-targeted SPL transcription factor［J］.The Plant Cell，2011，23（4）：1512-1522.
11	SCHWARZ S， GRANDE A V， BUJDOSO N，et al.The microRNA regulated SBP-box genes SPL9 and SPL15 control shoot maturation in Arabidopsis ［J］.Plant Molecular Biology，2008，67（1/2）：183-195.
12	SALINAS M， XING S P， H?HMANN S，et al.Genomic organization，phylogenetic comparison and differential expression of the SBP-box family of transcription factors in tomato［J］.Planta，2012，235（6）：1171-1184.
13	WANG S K， WU K， YUAN Q B，et al.Control of grain size，shape and quality by OsSPL16 in rice［J］.Nature Genetics，2012，44（8）：950-954.
14	WANG L， ZHANG Q F.Boosting rice yield by fine-tuning SPL gene expression［J］.Trends in Plant Science，2017，22（8）：643-646.
15	HOU H M， LI J， GAO M，et al.Genomic organization，phylogenetic comparison and differential expression of the SBP-box family genes in grape［J］.PLoS One，2013，8（3）：e59358.
16	HIRATSU K， MATSUI K， KOYAMA T，et al.Dominant repression of target genes by chimeric repressors that include the EAR motif，a repression domain，in Arabidopsis ［J］.The Plant Journal，2003，34（5）：733-739.
17	杨洋.白桦BpTCP7基因的功能研究［D］.哈尔滨：东北林业大学，2016.
17	YANG Y.Function analysis of BpTCP7 gene in Betula platyphylla ［D］.Harbin：Northeast Forestry University，2016.
18	YU N， YANG J C， YIN G T，et al.Genome-wide characterization of theSPL gene family involved in the age development of Jatropha curcas ［J］.BMC Genomics，2020，21（1）：368.
19	GAO J， PENG H， CHEN F B，et al.Genome-wide identification and characterization，phylogenetic comparison and expression profiles of SPL transcription factor family in B.juncea（Cruciferae）［J］.PLoS One，2019，14（11）：e0224704.
20	WANG J W， SCHWAB R， CZECH B，et al.Dual effects of miR156-targeted SPL genes and CYP78A5/KLUH on plastochron length and organ size in Arabidopsis thaliana ［J］.The Plant Cell，2008，20（5）：1231-1243.
21	WU G， POETHIG R S.Temporal regulation of shoot development in Arabidopsis thaliana by miR156 and its target SPL3 ［J］.Development，2006，133（18）：3539-3547.
22	SHIKATA M， KOYAMA T， MITSUDA N，et al. Arabidopsis SBP-box genes SPL10，SPL11 and SPL2 control morphological change in association with shoot maturation in the reproductive phase［J］.Plant and Cell Physiology，2009，50（12）：2133-2145.
23	WU G， PARK M Y， CONWAY S R，et al.The sequential action of miR156and miR172 regulates developmental timing in Arabidopsis ［J］.Cell，2009，138（4）：750-759.
24	LI L C， QIN G J， TSUGE T，et al. SPOROCYTELESS modulates YUCCA expression to regulate the development of lateral organs in Arabidopsis ［J］.New Phytologist，2008，179（3）：751-764.
25	LIU K Y， CAO J， YU K H，et al.Wheat TaSPL8 modulates leaf angle through auxin and brassinosteroid signaling［J］.Plant Physiology，2019，181（1）：179-194.
26	DU Y F， LIU L， LI M F，et al. UNBRANCHED3 regulates branching by modulating cytokinin biosynthesis and signaling in maize and rice［J］.New Phytologist，2017，214（2）：721-733.

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