Comparative Study On Anther Proteins Of Male Sterility Induced By Chemical Hybridizing Agent And CMS Gene In Wheat

Wheat (Triticum aestivumL.) has also obvious heterosis as other crops. The heterosis of wheat can be deeply explored and can be utilized to improving its quality and yield per unit of area sown. Now there are many important methods used for producing hybrid wheat seed to utilize the heterosis of wheat. The Chemical Hybridizing Agent System(CHAS) developed rapidly and had become a main way in the applying with producing in recent years. Because the male and female parent in CHAS can be selected freely and combined from the conventional wheat varieties as to increase the efficiency of hybrid breeding programs. SQ-1 is a new kind of CHA with self-knowledge property in china. SQ-1 could induce above 95% male sterility and has no side-effect on growth and development in wheat, and resolve the restriction of Chemical Hybridizing Agent System in our country.In general, the male sterility that induced by the chemical Hybridizing Agent belong to physiological male sterility. The cytoplasm male sterility system that controlled by male sterility gene, such as male sterility three-line, it was used that male sterility of Nucleo-Cytoplasmic interaction type to produce hybrid seeds, so it belong to hereditable male sterility. In order to reveal deeply the mechanism of wheat male sterility, this paper compared the difference from protein level between hereditable male sterility and physiological male sterility in same condition. We hope to discovery the similarities and differences in the course of pollen abortion, to provide theoretical basis for the mechanism of male sterility and to provide technics support for Hybrid Wheat in production practice.The anthers of isonucleus-allocytoplasmic materials of Xinong1376, ms(S)-Xinong1376, and ms(A)-Xinong1376 were used to investigate the protein expression profile in wheat. By utilization of SDS-PAGE and IEF/ SDS-PAGE, we compared and analyzed the specific of anthers protein. The main results obtained were as follows :1. The pollen abortion processes were investigated using aceto-carmine squash method and estimated the development stages under optical microscope. The results showed that the pollen of Xinong1376 are normal, and two male sterile lines have more pollen abortion in mononucleated prophase stage.2. Using two different ways to extract the protein from whaet anthers respectively, we of anthers protein at the same loading quantities. The proteins extracted by TCA/acetone strategy possess following strong points: the higher extraction rate, good quality, high purity, and the clear banding patterns. However, The proteins extracted by the other method cannot be relied in differential analysis of protein expression.3. The anthers proteins of Xinong1376, ms(S)-Xinong1376, and ms(A)-Xinong1376, including mononucleated prophase, binucleated stage, and trinucleated stage, was studied by polyacrylamide gelelectrophoresis technology. There are almost no difference in protein bands among the three materials. It might be explained by: (1) There have the same cell nuclear genetic background; (2) The pollen abortion is a very complex process. SDS-PAGE has so low resolution that cannot effectively proved the protein expression mechanism of pollen abortion.4. We investigate the protein expression profile in anthers of the isonucleus-allocytoplasmic materials: Xinong1376, ms(S)-Xinong1376, and ms(A)-Xinong1376 by two-dimensional electrophoresis. About 320 to 350 distinct protein spots were detected reproducibly. The results indicated that there were some difference in the protein quantity between the two sterile lines and the maintainer line. Several specific spots were present in the maps of two sterile lines, but were absent in the maintainer line. The two sterile lines had their own specific expression protein spots with different sterile mechanism. We compared the three maps and concluded that the sterile lines may inhibit related protein expression, and turned on some proteins related to male sterility, which disturbs the normal metabolism of substances and energy in anthers and leads to the occurrence of male-sterility.