Molecular Cloning and Functional Identification of a Pericarp- and Testa-Abundant Gene’s (AhN8DT-2) Promoter from Arachis hypogaea

Author:

Sharif Yasir12ORCID,Zhuang Yuhui12,Xie Wenpin1,Zhang Chong1,Chen Kun13,Deng Ye1,Chen Yuting1,Fu Huiwen13,Wang Lihui13,Chen Xiangyu14,Zhuang Weijian1ORCID,Chen Hua1

Affiliation:

1. College of Agriculture, Center of Legume Plant Genetics and System Biology, Institute of Oil Crops, Fujian Agriculture and Forestry University (FAFU), Fuzhou 350002, China

2. College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China

3. College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002, China

4. Crops Research Institute, Fujian Academy of Agricultural Science, Fuzhou 350013, China

Abstract

Cultivated peanut (Arachis hypogaea L.) is a key oil- and protein-providing legume crop of the world. It is full of nutrients, and its nutrient profile is comparable to that of other nuts. Peanut is a unique plant as it showcases a pegging phenomenon, producing flowers above ground, and after fertilization, the developing peg enters the soil and produces seeds underground. This geocarpic nature of peanut exposes its seeds to soil pathogens. Peanut seeds are protected by an inedible pericarp and testa. The pericarp- and testa-specific promoters can be effectively used to improve the seed defense. We identified a pericarp- and testa-abundant expression gene (AhN8DT-2) from available transcriptome expression data, whose tissue-specific expression was further confirmed by the qRT-PCR. The 1827bp promoter sequence was used to construct the expression vector using the pMDC164 vector for further analysis. Quantitative expression of the GUS gene in transgenic Arabidopsis plants showed its high expression in the pericarp. GUS staining showed a deep blue color in the pericarp and testa. Cryostat sectioning of stained Arabidopsis seeds showed that expression is only limited to seed coat (testa), and staining was not present in cotyledons and embryos. GUS staining was not detected in any other tissues, including seedlings, leaves, stems, and roots, except for some staining in flowers. Under different phytohormones, this promoter did not show an increase in expression level. These results indicated that the AhN8DT-2 promoter drives GUS gene expression in a pericarp- and testa-specific manner. The identified promoter can be utilized to drive disease resistance genes, specifically in the pericarp and testa, enhancing peanut seed defense against soil-borne pathogens. This approach has broader implications for improving the resilience of peanut crops and other legumes, contributing to sustainable agricultural practices and food security.

Funder

National Natural Science Foundation of China

Publisher

MDPI AG

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