The Construction Of Mutant Populations Induced By Physical And Chemical Mutagen And Screening Of Iron Uptake Mutants In Azuki Bean(Vigna Angularisi)

Azuki bean(Vigna angularisi) originated from China is traditionally cultivated crop. Total planting area and yield of azuki bean in china is the maximum in the world. Mutagenesis have been widely applied in breeding, genetics and gene functional research. We have firstly developed a large collection of mutants by mutagenizing azuki bean (Vigna angularisi) cultivar Jingnong 6 with EMS (Ethyl Methan Sulfonate), electron beam and 60Co y-ray. The construction of saturated mutant population is very important and indispensable for gene mapping and cloning, functional genomics research in azuki beanCultivar Jingnong 6 of azuki bean was induced with 0.5%,0.9%,1.4% concentration EMS for 12h and 24h,400Gy 60Co y-ray and 100Gy,300Gy,600Gy electron beam. Progenies from M1 to M4 were investigated. Mutants were screened, then their trait genetic stability was analyzed. New leaf yellowing mutants were specially identified in Fe stress Hoagland hydroponic solution. Agronomic traits of the mutants were evaluated. Mutant populations and a database have been constructed and developed.1. A total of 11240 seeds of cultivar Jingnong 6 were treated with EMS,400Gy 60Co y-ray and electron beam. EMS-induced 937 M3 lines and 650 M4 family lines, and 934 M2 lines and 359 M3 family lines radiated with 400Gy 60Co y-ray and Electron beam were generated.1490 mutants have been created. Mutation involves in plant height, plant architecture, leaf shape and color, leaf size, seed shape and color, seed size, pod color, branching type and number, bushy leaf or branch, sprawl, definite growth, sterile, early mature and late mature, flowering time and nutrient uptake.2. The different mutant traits such as definite growth, pale green leaves, yellowing lesf, sword leaf, kidney leaf, small heart-shaped leaf, needle leaf, smooth leaf, cluster leaf,, high plant, dwarf, early or late mature, plant architecture, black pod, dark brown pod, brown pods, yellow and white seed, samll or large seed size can be stably inherited for three years.3. The broader mutantions are induced by EMS. The cost-effective concentration is 0.5% or 0.9% to treat for 24h. More seed color mutants including cream-colored and light red, pod color mutants such as brown, dark brown and black are produced by o.5% EMS. Needle leaf, sword, kidney-shaped and small heart-shaped leaf mutants are obtained by 0.9% EMS treatment. The radiational dose of Electron beam are 100,300,600Gy, the percentage of mutants are 4.09%,3.64%,2.22%. The frequency of mutants treated by 400Gyγ-ray is 7.23%.4. New leaf yellowing mutants were tested in stress of iron deficiency, normal iron, high iron concentrations of Hoagland hydroponic solution.1 iron-deficient stress sensitive mutant and 1 insensitive mutant have been identified. Research shows that the reductase activity of normal plants,low Fe-sensitive plants and Low-Fe tolerance mutant in iron-deficiency stress were higher than the activity in normal iron environment. The reductase activity of low Fe-sensitive mutant in iron-deficiency stress was the highest, control was in the center, Low-Fe tolerance mutant was the lowest. Fe,Mn,Zn,Ca,Mg,K and P element contents and change of low-Fe-sensitive mutant in root were consistent with the control. But the Fe element content of low-Fe-sensitive mutant in leaves was significantly reduced than the control, it indicated low Fe-sensitive mutants developed into yellow because its shoot transport which may be a new type mutant. Ability of iron uptake and transport of low-Fe tolerance mutant were higher than control. Under iron-deficiency stress, the iron content in its leaves was higher than the control and Fe-sensitive mutant, the rest element content of low-Fe tolerance mutant had similar trend with the control. The mutants will be researched in genetic analysis, gene mapping and cloning.5. Comprehensive evaluation of the mutant agronomic traits showed that light green leaves, small leaf, compact and upright plant, black pod, light red seed, yellow and white seed, small or large-sized seed were superior to the control. The agronomic traits of yellowing plant, sword leaf, small heart-shaped leaf, needle leaf, smooth leaf surface, cluster leaf,, dwarf plant, brown pod mutants were inferior to control, their yield decreased significantly. The mutant agronomic traits of dark green plant, kidney leaf, loose plant, high plant, dark brown pod, dark red seed were similar to the control.10 excellent lines have been selected for comparative test and breeding.The mutants of different genetic variation are suitable for both forward and reverse genetics. The mutant populations are very useful to genetic analysis of gene, gene mapping and cloning, and further the research on functional genomics. These mutants will be very useful to serve for azuki bean improvement.