Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (9): 1723-1732.DOI: 10.3969/j.issn.1000-2561.2020.09.001

• Omics & Biotechnology •     Next Articles

Cloning, Physical Location and Expression Analysis of HbGRF in Hevea brasiliensis

ZHANG Yuhang1,2,*,PAN Ranran1,*,LI Fei1,TAO Zhiqiang1,2,WANG Ying1,GAO Heqiong1,ZHUANG Nansheng1,**()   

  1. 1. College of Tropical Crops, Hainan University, Haikou, Hainan 570228, China
    2. Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
  • Received:2019-09-09 Revised:2019-12-23 Online:2020-09-25 Published:2020-10-16
  • Contact: ZHANG Yuhang,PAN Ranran,ZHUANG Nansheng

Abstract:

Growth-regulating factor (GRF) is a plant-specific transcription factor. The transcription factor family has a large number of members and plays an important role in plant growth and development. In this study, the excellent clone ‘Reyan 7-33-97’ was used as the experimental material, and the HbGRF genes were cloned by RT-PCR. The bioinformatics method was used to study its gene length, physicochemical properties, basic structure and evolutionary relationship. The physical localization analysis was carried out by in situ PCR and fluorescence in situ hybridization. The expression pattern of HbGRF genes in rubber trees was analyzed by qRT-PCR. Three GRF genes were cloned into the female flowers of rubber trees, named HbGRF1, HbGRF2 and HbGRF3, respectively. The protein length was 609 aa, 384 aa and 494 aa with molecular weight of 65.663 kDa, 41.188 kDa and 52.858 kDa, respectively, and they were all unstable protein. All the three members had the complete characteristic domains WRC and QLQ of GRF gene family. Evolutionary analysis showed that the three genes belonged to three different groups. HbGRF1, HbGRF2 and HbGRF3 were located on the long arm of chromosome 11, the short arm of chromosome 5 and the long arm of chromosome 9, and the average percentage distance between the signal site and the centromere was 77.65, 42.66 and 65.27, respectively. The expression results showed that the three HbGRF genes in rubber trees were highly expressed in the vigorous growth tissues like stem tips and female flowers. Under the treatment of exogenous GA and ABA, the expression levels of the three genes increased first, and the expression levels at 48 h were basically the same as those under the initial conditions. The three genes had obvious expression specificity in stem tips and female flowers, which may play important roles in flower bud differentiation and female flower development. This study would provide a theoretical basis and materials for further exploration of the function of HbGRF, and provide molecular cytology basis for accurate breeding of rubber trees.

Key words: Hevea brasiliensis, GRF gene family, gene mapping, expression analysis

CLC Number: