| In this report, methods of introducing theγ-tmt and bar gene into peanut cultivars(Fenghua1,Baisha1016,Fenghua3 and 79266) with Pollen tube pathway and In Planta transformation were studied. The setting rate and frequency of plantlets of two transgenic methods were compared. The appropriate method of screening transformed gene plants was discussed. The three main pathogenic fungi of peanut were isolated by tissue isolating method and the affecting factors on the mycelium and sporulation of Cercospora arachidicola Hori fungi were investigated. The pathogenic power was also investigated. The DNA from Chinese scholar tree and A.glabrate was introduced into two peanut cultivar 79266 and Baisha1016 by the exotic DNA introduction through pollen tube pathway. Seven varieties of leafspot disease-resistance were selected from the varieties. Resistance to peanut leafspot diseases of these varieties was identified. Quality traits of these varieties were compared. And the SSR polymorphic analysis was also investigated. The major results are abstracted as follows:1. Studies on the genetic transformation with pollen tube pathway and In Planta transformation techniquesWith the Pollen tube pathway and In Planta transformation, the transgenic operation was done. The setting rate and frequency of plantlets by two transgenic methods were compared. It showed that setting rate and frequency of plantlets with using pollen tube pathway were higher than that with using In Planta transformation. Appropriate concentration of PPT by screening leaves in field to select transgenic plants was 60 mg/L.2. The isolation of pathogenic fungi in peanut early leafspot disease, late leafspot disease and web blotch disease The pathogenic fungi in peanut early leaf spot disease, late leaf spot disease and web blotch disease were isolated by tissue isolating method. The affecting factors on the mycelium and sporulation of pathogenic fungi in peanut early leaf spot disease were investigated. And the pathogenic power was also investigated by leaf in vitro inoculation and spraying total plant with sporule suspension. The result showed that the pathogenic fungi could grow and sporulate between 15~30℃, and the best suitable temperature is 25℃. In addition, using black light could accelerate sporulation. The pathogenesis experiments showed that this fungus could infect peanut by two different methods, but the blotch of disease leaves was different.3. Identification of leafspot-disease -resistance varieties(1) Compare of leafspot-disease mean grade between varieties and acceptors Seven varieties were identified resistance to peanut early leafspot diseaseand web blotch disease. The results indicated that variety 3 obtained from Chinese scholar-tree DNA intruducing peanut cultivar 79266 had the highest resistance to peanut early leafspot disease, and its mean grade was only 2.72. Variety 4 had the second highest resistance to peanut early leafspot disease, and its mean grade was 2.82. These two varieties'mean grade decreased significantly compared to the acceptor. The mean grade of variety 1,2 and 5 was 3.16,3.00 and 3.25 respectively. These varieties'mean grade decreased. Two varieties obtained from A.glabrate DNA intruducing peanut cultivar Baisha1016 showed significantly higher resistance to peanut early leafspot disease. The mean grade of variety 6 and variety 7 was 3.36 and 2.91 respectively, while the Baisha1016'mean grade was 4.15.The results also indicated that variety 2 had the highest resistance to web blotch diseases, and its mean grade was only 3.11. Variety 3 had the second highest resistance to web blotch diseases, and its mean grade was 3.19. While the acceptor'mean grade was 4.56. The mean grade of variety 1,4 and 5 was 3.90,3.62 and 3.65 respectively. All of the five varieties'mean grade decreased significantly compared to the acceptor. Variety 6 and variety 7 also had higher resistance to web blotch disease compared to Baisha1016. The two varieties'mean grade was 4.10 and 3.68, while the Baisha1016'mean grade was 5.62. These two varieties'mean grade decreased significantly compared to the acceptor.(2) Compare of AUDPC between varieties and acceptorsThe AUDPC of 79266 was 205.33, while the varieties'AUDPC was 112.00, 116.67, 133.00 and 145.83 respectively from variety 1 to 5. The AUDPC of Baisha1016 was 240.33, while the varieties'AUDPC was 170.33 and 177.33 respectively. The leafspot disease of varieties decreased significantly compared to the acceptors.(3) Compare of agronomic characters between varieties and acceptorsIt showed that most of agronomic characters changed because of the exotic DNA introduction. Plant height of 79266 was 63cm, while plant height of variety 1 and variety 4 was only 39cm and 54cm. They all decreased significantly compared to acceptor. Length of the first branches of 79266 was 68cm, while the length of the first branches of variety 4 was only 52cm. There was 15 pods in each plant of 79266, while the pods in each plant of 1,2 and 3 were 46,30 and 31 respectively. They all increased compared to the acceptor. 100-pods weight and Pod weight of one plant of 79266 was 271g and 25.93g respectively. 100-pods weight of variety 1,3,4 and 5 was 133g,208g,246g and 237g respectively. They decreased compared to the acceptor. Pod weight of one plant of variety 2,3,4 and 5 was 14.32g,21.99g,12.32g and 22.71g respectively. They all decreased. 100-pods weight and pod weight of one plant of Baisha1016 was74g and 20.76g respectively. 100-pods weight of variety 6 was 66g, it decreased compared to the acceptor. While that of variety 7 was 89g, it increased compared to Baisha1016. Pod weight of one plant of variety 6 was 15.33g, it decreased compared to the acceptor. The results also indicated that there was no significant correlation between resistance level with agronomic characters.(4) Compare of quality traits between varieties and acceptorsThe protein content, crude lipid content, soluble total sugar content and O/L of 79266 was 24.69%,54.1%,3.54% and 2.16 respectively. While the protein content of variety 2 and variety 4 was 26.53% and 26.13%. They increased compared to the acceptor. The protein content of variety 1 and variety 3 was 23.75% and 23.00%. They decreased compared to the acceptor. The crude lipid content of variety 2 and variety 3 was 49.43% and 52.03%. They decreased compared to the acceptor. The soluble total sugar content of variety 1,2 and 3 was 3.81%,4.25% and 4.29% respectively. They all increased. While that of variety 4 was only 2.86%, and it decreased compared to the acceptor. The O/L of variety 2 and 4 was 1.79 and 1.26, and they decreased. While that of variety 3 was 2.38, and it increased compared to the acceptor. The protein content, crude lipid content, soluble total sugar content and O/L of Baisha1016 was 27.32%,53.73%,3.82% and 1.23 respectively. While the protein content and crude lipid content of variety 6 was 24.82% and 51.13%, and they all decreased compared to the acceptor. The soluble total sugar content and O/L of variety 6 was 4.60% and 2.04, and they increased compared to the acceptor of Baisha1016. The soluble total sugar content and O/L of variety 7 was 5.24% and 2.32, and they increased compared to the acceptor.(5) SSR polymorphic analysis of the varieties and acceptorsSSR polymorphic analysis showed that there was 19,90,18,19,20,17 and 15 primers that can produce bands in variety 1 to variety 7. And there was 6,7,5,6,4,4 and 3 polymorphic primers in seven varieties respectively. Using the polymorphic primers it can produce 22,24,22,23,23,24 and 24 bands respectively between the varieties and acceptors. And there was 9,8,6,7,5,6 and 5 polymorphic bands respectively between the varieties and acceptors. Variet 1 has the most difference in molecular level compared to its acceptor.Variety 5 and variety 7 had the least difference in molecular level compared to their acceptor.
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