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使用三步培养法对粳绿稻品系 810 10 4系进行花药培养。以N6 + 2 ,4 D 1.0mg/L +NAA 3.0mg/L +KT 1.0mg/L +椰乳 10 0mL/L +麦芽糖 5 0 g/L +Na2 SiO36 0mg/L +琼脂粉 7g/L(pH5 .8)作为愈伤组织诱导培养基 ,MS +BA 1.0mg/L +KT 1.0mg/L +NAA 0 .5mg/L +蔗糖 30 g/L +琼脂粉 7g/L(pH5 .8)作为分化培养基。愈伤组织诱导率为 3.33% ,绿苗分化率为 11.4 9%。获得绿色花培再生植株 18丛 ,分为 39个单株系。经田间形态观察和染色体计数鉴定 ,其中有 33个株系为单倍体 ,6个株系为二倍体。 6个二倍体株系中 ,有 2个株系的粒色表现为纯合体 ,有 4个株系的粒色表现为杂合体。 4个杂合体株系中 ,有一个显现出花叶性状。对花药培养过程中的形态变化观察结果表明 ,在三步培养法中 ,愈伤组织分化形成胚状体和不定芽 ,其中以形成胚状体为主 ;在愈伤组织直径约为 1mm时转入分化培养基为佳
The three-step culture method was used for anther culture of japonica rice line 810104. The optimal conditions were as follows: N6 + 2, 4 D 1.0 mg / L + NAA 3.0 mg / L + KT 1.0 mg / L + coconut milk 10 0 mL / L + maltose 50 g / L + Na2SiO36 0 mg / L + agar powder 7 g / pH5.8) as callus induction medium, MS + BA 1.0 mg / L + KT 1.0 mg / L + NAA 0.5 mg / L + sucrose 30 g / L + agar powder 7 g / L Differentiation medium. The callus induction rate was 3.33% and the green plantlet differentiation rate was 11.49%. Eighteen clumps of regenerated plants were obtained and divided into 39 lines. After field morphological observation and chromosome count identification, 33 of them were haploid and 6 were diploid. Among the 6 diploid lines, 2 of the lines showed homogeneity in their color, and 4 lines showed heterozygosity. One of the four hybrids showed leafy traits. Morphological changes in the process of anther culture observed results showed that in the three-step culture method, the callus differentiated to form embryoid body and adventitious buds, of which the formation of embryoid body; callus diameter of about 1mm Differentiation medium is better