Cloning and Expression Activity Analysis of Rubisco Small Subunit Gene Promoter in Populus × xiaohei

doi:10.7525/j.issn.1673-5102.2024.04.014

Abstract

Abstract:

Ribulose-1，5-diphosphate carboxylase/oxygenase（Rubisco） is a key enzyme in photosynthesis， in which small subunits（rbcS） are encoded by nuclear genes and mainly expressed in leaves. In this study， the Rubisco small subunit genes PxrbcS1 and PxrbcS2， which are highly expressed in Populus × xiaohei’s leaves， were determined by RT-qPCR technology， and their upstream promoters of 2 240 and 2 174 bp were cloned respectively. The results of promoter element analysis showed that the promoters of PxrbcS1 and PxrbcS2 had multiple elements related to light induced expression， including G-box， MRE and Box4 elements. Plant expression vectors of pBI-121-pPxrbcS1：：GUS and pBI-121-pPxrbcS2：：GUS were constructed and genetically transformed into 84K poplar（P. alba×P. glandulosa）. GUS histochemical staining and qPCR expression analysis showed that the promoters of PxrbcS1 and PxrbcS2 could drive the GUS gene to express in 84K poplar leaves with high specificity. In conclusion， the above results showed that PxrbcS1 and PxrbcS2 promoters with high leaf expression activity were successfully cloned from P.×xiaohei， and the promoters might be applied to the study of gene functions related to plant photosynthesis and genetic operations to improve photosynthesis in plants， including poplars.

Key words: Poplar, rubisco small subunit promoter, cloning, regulatory element, genetic transformation, GUS staining

CLC Number:

Jiayu CAO, Lina CAO, Qiaoyi ZHANG, Shuang ZHANG, Zhibao HU, Tangrui ZHAO, Zhiru XU, Chunming LI, Xiankui QUAN, Guanjun LIU. Cloning and Expression Activity Analysis of Rubisco Small Subunit Gene Promoter in Populus × xiaohei[J]. Bulletin of Botanical Research, 2024, 44(4): 625-633.

Figures/Tables 9

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元件 Elements	序列 Sequence	功能 Function
Box 4	ATTAAT	参与光响应的顺式作用元件Part of a conserved DNA module involved in light responsiveness
TCT-motif	TCTTAC	参与光响应的顺式作用元件Part of a light responsive element
G-box	TACGTG	参与光响应的顺式作用元件Cis-acting regulatory element involved in light responsiveness
ATC-motif	AGTAATCT	参与光响应的顺式作用元件Part of a conserved DNA module involved in light responsiveness
ABRE	ACGTG	参与脱落酸反应的顺式作用元件Cis-acting element involved in the abscisic acid responsiveness
TC-rich repeats	ATTCTCTAAC	参与防御和应激反应的顺式作用元件Cis-acting element involved in defense and stress responsiveness
CAT-box	GCCACT	参与分生组织调控的顺式作用元件Cis-acting regulatory element related to meristem expression
MRE	AACCTAA	参与光响应的myb结合位点myb binding site involved in light responsiveness
TCA-element	CCATCTTTTT	参与水杨酸反应的顺式作用元件Cis-acting element involved in salicylic acid responsiveness
MBS	CAACTG	参与干旱诱导的myb结合位点myb binding site involved in drought-inducibility

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