![]() Using the CRISPR/Cas9 system to create targeted mutations in MS1, a double displacement code mutation was introduced into MS1, which resulted in complete male sterility in wheat varieties Fielder and Gladius. The results showed that the MS1 gene in Brassica crops belonged to the PHD finger family, and its highly conserved sequence was involved in the regulation of pollen development and maturation. Using the MS1 gene in Arabidopsis thaliana as a reference sequence, bioinformatic methods were applied to predict and analyze the amino acid sequences encoded by Brassica crops such as rape, Chinese cabbage, and cabbage. The CRISPR/Cas9 system was used to develop male-sterile lines without transgenic maize and obtain stable genetic lines. It is speculated that the Zmms1 gene, along with Osms1, may control fertility. The homologous Zmms1 gene in maize was cloned from the Osms1 gene in rice by homologous cloning, and the results of the bioinformatics analysis showed that both Zmms1 and Osms1 were very similar to the members of the PHD domain family. The PTC1 gene in rice was found to be homologous to the MS1 gene in Arabidopsis, which also encodes a PHD domain and shows similar functions in regulating the development of villus cells and the pollen wall. They cause pollen abortion, and thus, can be used in production. PHD is closely related to the composition of the pollen wall and the tapetum development and does not affect other floral organs. Among them, the MS1 gene, which has been proven to be a key gene regulating pollen formation, is mainly expressed when microspores are released from the tetrad and encodes a plant homeodomain (PHD)-containing nuclear protein. To date, 13 genes associated with male sterility have been cloned from Arabidopsis and maize. The sterile males are used to produce hybrids that can save artificial castration, reduce seed production costs, and improve seed purity. It is a genetic phenomenon defined by the inability of plants to produce normal anthers, pollens, or male gametes during sexual reproduction, while the pistil functions normally. Male sterility is an important agronomic trait in studies of crop heterosis utilization. The results showed that the Ntms1 gene plays an important role in regulating fertility in tobacco. The malondialdehyde content in the mutant plants was significantly higher than that in the wild-type plants, while self-fertility was significantly lower in the mutant plants. The anthers did not develop fully and had few viable pollen grains the tapetum and the anther wall had developed abnormally, and the anther chamber was severely squeezed. Compared to the wild-type plants, the filaments of the Ntms1 knockout mutant plants were shorter, and the stamen was shorter than the pistil. It is speculated that the Ntms1 gene had the same function as the Cams1 gene in controlling male sterility. The results showed that the sequences of Ntms1 and Cams1 were 85.25% similar, and plant homeodomains were found in both genes the physical and chemical properties were also very similar. The protein structure and physicochemical properties of the two genes were determined by bioinformatics analysis, and the function of the Ntms1 gene was verified by the CRISPR/Cas9 system. ms1) gene sequence of male-sterility genes in pepper. ms1) gene was cloned from the tobacco variety K326 by homologous cloning based on the Cams1 ( Capsicum annuum L. To broaden the sources of male sterility in tobacco, the Ntms1 (Nicotiana tabacum L. The sterile line is the basis of crop heterosis utilization.
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