| Alien additional line is a significant bridge material in the process of establishing alien substitution lines and translocation lines, with an important value in theoretical research and breeding practice. The accurate identification of alien chromosome and its genetic analysis are the precondition for the effective utilization of alien addition line. Among the various identification methods, the polymorphic In Del markers developed based on the data of genome re-sequencing could provide the technical guarantee for the accurate identification of alien additional line. The genetic analysis of alien chromosome includes the transmission rate and the size of transmission fragments, which determines chromosomal number and composition of the derived offspring from alien additional line In order to innovate the genetic germplasm of Chinese cabbage(Brassica rapa L. ssp. pekinensis, AA) and enrich its genetic background, the studies of transferring the valuable genes of head cabbage into Chinese cabbage were carried out in recent years, and a set of Chinese cabbage-cabbage additional lines and part of disomic additional lines were obtained. But the genetic analysis of alien chromosome in the derivative progenies has not been systematically studied.In this research, In Del markers and cytological method were applied to identify chromosomal number and organization of the self progenies and microspore regeneration plants of Chinese cabbage-head cabbage No.1 chromosome disomic additional line(AC1D) and the radiant mutant progenies of Chinese cabbage-head cabbage No.1 chromosome monosomic additional line(AC1M). Meanwhile, the characteristics of two translocation lines were studied. The main results are as follows:1. With Chinese cabbage ‘85-1’ and head cabbage ‘11-1’ as the control materials, 6~8 In Del makers that evenly distributing on each of 9 cabbage chromosomes were applied to amplify two plants with chromosome number of 21 and 22, respectively, and from the self progenies of Chinese cabbage-head cabbage AC1 D. The results showed that Chinese cabbage-head cabbage No.1 chromosome additional line was added cabbage chromosome C03 and infiltrated with some fragment of C07 and C05 chromosomes. Further, 67 polymorphic In Del markers distributed along cabbage chromosome C03 were screened for the two plants. The specific bands of cabbage were detected by 52 In Del markers on the plant with 21 chromosomes and 40 In Del markers on the plant with 22 chromosomes. The two plants were inferred as Chinese cabbage-head cabbage No.1 chromosome monosomic and disomic additional lines, respectively.2. The self progenies of Chinese cabbage-head cabbage AC1 D were identified with the PCR technique of 67 polymorphic In Del markers from C03 chromosome and cytological method. The result showed that 3 individual plants with 22 chromosomes were inferred as Chinese cabbage-head cabbage No.1 chromosome disomic additional lines, 6 individual plants with 21 chromosomes were inferred as Chinese cabbage-head cabbage No.1 chromosome monosomic additional lines, and 29 individual plants with 20 chromosomes were inferred as Chinese cabbage-head cabbage No.1 chromosome translocation lines. In addition, 3 individual plants with 22 chromosomes and 2 individual plants with 21 chromosomes were inferred as Chinese cabbage double trisomics and trisomics, respectively.3. Twenty three individual plants from the self progenies of Chinese cabbage-head cabbage AC1 D were isolated microspore culture. Fourteen individual plants produced embryos, and 9 of them obtained microspore plants. The embryo rate of these plants was 0.1~4.2 embryos/bud, and the plant regenerated rate was 7.1%~100%. In total 139 isolated plants were obtained. Using the specific makers carried by their parental plant 7, 8, 24 and 47, their microspore plants and the self two-generation progenies were identified. In the microspore plants 3, 2, 0, and 1 plants carried with the specific makers of cabbage were identified, respectively, and the ratio of the identified plants to the total plants was 12%, 66.7%, 0 and 20%, respectively. In the self two-generation progenies 3, 11, 18 and 26 plants carried with cabbage specific makers of were identified, respectively, and the ratio of the identified plants to the total plants was 13%, 28.2%, 23.1% and 20%, respectively. In all, 9 plants carried with 1 cabbage specific In Del maker and 13 plants carried with 3~9 continuous cabbage specific In Del maker were obtained.4.The radiant mutant of backcross progenies from Chinese cabbage-head cabbage No.1 chromosome monosomic additional line were identified using 67 polymorphic In Del markers of chromosome C03. And 15 of 68 backcross progenies were detected the specific bands of head cabbage. Among 19 plants, 1 plant(BC-3) was detected 12 specific In Del makers, 1 plant(BC-2) was detected 2 specific In Del makers, the other 13 plants were detected only 1 specific In Del makers. The number of chromosomes in 6 plants(BC-1, BC-2, BC-3, BC-8, BC-10 and BC-15) was identified, showing that the BC-3 individual plant has 21 chromosomes, which was inferred as Chinese cabbage-head cabbage No.1 chromosome monosomic additional line. And the other 5 plants all had 20 chromosomes, which were inferred as Chinese cabbage-head cabbage No.1 chromosome translocation lines. The transmission rate of the specific makers carried by the 6 individual plants was further identified. The specific bands of cabbage were detected only in the self offspring of BC-1 and BC-15, the ratio of the identified plants to the total plants were 1.5% and 13.9%, respectively. While no specific bands of cabbage were detected in the self progenies of the other 4 plants.5.The characters of two Chinese cabbage-head cabbage No.1 chromosome translocation lines were investigated in the period of vegetable growth, and the nutrient content were measured in the harvest stage. The result showed that there was differences in leaf and plant morphology bewteen the self progenies of two translocation lines and diploid Chinese cabbage ’85-1’. The nutritional indicators detection showed that there was significant differences in the mean Vc contents, the mean soluble sugar content and the mean dry matter content among the test materials, while the mean cellulose content were not significantly different among these plants. The mineral elements detection showed that there was significant differences in the mean Ca content and the five trace element among the test material. Key words: Chinese cabbage; Head cabbage; Alien additional line; Microspore culture;... |
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