Bioinformatics Analysis and Functional Verification of ZjIAA9 in Ziziphus jujuba Mill.

doi:10.7668/hbnxb.20195979

Abstract

Abstract:

The members of the Aux/IAA gene family encode proteins that regulate various developmental processes in plants,such as embryogenesis and seed development,by modulating the auxin (IAA) signal transduction pathway.To explore the role of Aux/IAA family genes in jujube embryo development,this study performed bioinformatics analysis and functional verification on the key genes of the Aux/IAA family that affect jujube embryo development,using transcriptome data of Huizao (a jujube variety with high embryo fertility) and Kongfusucui (a jujube variety with low embryo fertility).Results showed that an Aux/IAA family gene sequence with a length of 1 731 bp was obtained via gene cloning,which encoded 362 amino acids.Protein structure prediction indicated that its secondary structure was mainly composed of random coil and that it had a typical conserved domain of the IAA9 protein;thus,it was named ZjIAA9.Homology analysis revealed that ZjIAA9 was closely related to RGQ29_009000 in Quercus rubra and CFP56_014209 in Quercus suber.Transgenic Micro-Tom tomato plants were obtained via Agrobacterium-mediated leaf disc transformation.Determination of auxin content in transgenic Micro-Tom tomato plants showed that the IAA content in the pulp of transgenic plants was higher than that in non-transgenic plants,indicating that ZjIAA9 may regulate auxin biosynthesis.Compared with wild-type plants,transgenic plants showed a seedless or few-seeded phenotype,suggesting that jujube ZjIAA9 may be involved in regulating jujube embryo development.

Key words: Jujube, Embryo development, ZjIAA9, Bioinformatics analysis, Functional verification

CLC Number:

S665.1

YANG Mengfan, MA Qing, REN Haiyan, WANG Yongkang, ZHAO Ailing, XUE Xiaofang, SU Wanlong, SHI Meijuan, LIU Li, LI Yi. Bioinformatics Analysis and Functional Verification of ZjIAA9 in Ziziphus jujuba Mill.[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(6): 77-84. doi: 10.7668/hbnxb.20195979.

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http://www.hbnxb.net/EN/Y2025/V40/I6/77

Figures/Tables 10

Fig.1 RNA quality analysis 1—2.Embryo sac RNA of Huizao;3—4.Embryo sac RNA of Kongfusucui.

Fig.2 The expression level of ZjIAA9 in jujube embryos Different lowercase letters indicated significant differences between different treatments (P<0.05).The same as Fig.9.

Fig.3 The PCR product of the ZjIAA9 gene was detected by gel electrophoresis M.DNA Marker;1.PCR amplification product of ZjIAA9.

Fig.4 Bioinformatics analysis of ZjIAA9 A.Analysis of the hydrophobicity and hydrophilicity of the ZjIAA9 protein;B.Prediction of the structural domains of the ZjIAA9 protein; C.Prediction of the secondary structure of the ZjIAA9 protein;D.Prediction of the tertiary structure of the ZjIAA9 protein.

Fig.5 Multiple sequence alignment analysis of ZJIAA9 with its homologs from other species

Fig.6 Flow diagram of the transformation process of Micro-Tom cherry tomatoes A.Sowing;B.Induction of callus and adventitious buds from tomato leaves;C.Hygromycin resistance screening of adventitious buds;D.Induction of rooting.

Fig.7 PCR identification of transgenic plants M.2 000 bp DNA Marker;1—11.Transgenic plants; B.Blank control;N.Negative control;P.Positive control.

Fig.8 The influence of ZjIAA9 on the development of the seed kernels of transgenic cherry tomatoes

Fig.9 Analysis of the seed-to-fruit ratio between transgenic and non-transgenic cherry tomatoes

Fig.10 Comparison of IAA contents and relative gene expression levels between non-transgenic and transgenic cherry tomatoes IR.Control group kernel;N.Control group pulp;1—10.Pulp from ZjIAA9-transformed positive plants.Uppercase letters represent comparisons between the IR group and other groups; lowercase letters represent comparisons involving the N-10 group. Different letters indicate significant differences(P<0.05).

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