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薄壳山核桃嫁接愈合过程中CiMYB46基因的克隆与表达分析(PDF)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2019年05期
Page:
156-162
Column:
综合述评
publishdate:
2019-09-20

Article Info:/Info

Title:
Cloning and expression analysis of CiMYB46 in the graft healing process of Carya illinoinensis
Article ID:
1000-2006(2019)05-0156-07
Author(s):
MO Zhenghai1 2 LI Fengda1 SU Wenchuan1 CAO Fan1 PENG Fangren1* LI Yongrong3
(1.Co-Innovation Center for the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China; 2. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210014, China; 3. Green Universe Pecan Science and Technology Co., Ltd., Nanjing 210007, China)
Keywords:
Carya illinoinensis grafting gene cloning CiMYB46 MYB transcription factor
Classification number :
Q943.2; S722
DOI:
10.3969/j.issn.1000-2006.201811041
Document Code:
A
Abstract:
【Objective】 To explore the molecular mechanism of graft healing in pecan, we cloned the CiMYB46 gene and analyzed its expression pattern and cis-elements. 【Method】We extracted RNA from graft unions of pecan plants at different developmental stages and designed primers, based on our previous transcriptome results, to clone the desired gene by RT-PCR. In addition, genomic DNA was used as a template for PCR amplification and cloning of the promoter area of the desired gene.【Result】The amplified fragment obtained by RT-PCR was cloned and sequenced. It contained an open reading frame of 969 bp that started with the initiation codon, ATG and ended with the termination codon, TAA, encoding a polypeptide of 322 amino acids. The deduced amino acid sequence of the cloned gene contained a MYB domain and was closely clustered with Arabidopsis AtMYB46. Thus, we designated our sequence as CiMYB46. Real-time PCR analyses demonstrated that CiMYB46 was highly expressed during the vascular formation stage of graft union development in pecan and had a similar expression profile to that of some functional genes involved in secondary cell wall synthesis. A 1 070 bp promoter region upstream of the initiation codon of CiMYB46 was obtained through PCR and cloning, using genomic DNA as a template. PlantCARE analysis revealed that the promoter region of CiMYB46 contained two basic cis-acting elements, a CAAT box and a TATA-box, drought responsive elements, and hormone responsive elements, including ABA, Me-JA, GA, and SA. 【Conclusion】CiMYB46 might be associated with vascular bundle formation during the graft healing process of pecan. It may be induced by GA.

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Last Update: 2019-10-08