诸葛菜小孢子培养及其单倍体减数分裂染色体配对观察

诸葛菜是一种极有价值的观赏、蔬菜、饲料和油料作物种质资源。为建立诸葛菜小孢子胚状体诱导再生植株技术,并为诸葛菜染色体组的起源与进化研究提供相关数据资料,本研究通过对诸葛菜游离小孢子的培养,研究了热激培养时间和活性炭浓度对胚状体产量的影响,并采用常规压片法对诸葛菜单倍体减数分裂染色体配对行为进行了观察。结果表明,添加活性炭和热激培养对胚状体诱导是必需的。在直径6 cm培养皿中培养4 mL密度为1花蕾花粉mL?1的小孢子NLN悬液时,每皿添加1mg活性炭和32℃热激3d的培养条件下子叶形胚状体和总胚状体产量最高,分别为每花蕾0.92±0.18个和1.32±0.25个。子叶形胚状体在1/2 MS培养基上萌发率为27.73%。花粉植株中自然加倍率为25%,加倍植株染色体数为24,单倍体植株染色体数为12。诸葛菜单倍体减数分裂染色体的平均配对构型为n=12=6.352I+2.008II+0.384III+0.12IV,具有二价体及三价体和四价体的细胞比例高达96%,少量细胞的12条染色体联会形成3个四价体,说明诸葛菜很可能是起源于染色体基数x=3的同源八倍体。本试验结果对于诸葛菜新材料新品种选育...

Microspore culture and observations on meiotic chromosome pairing of the haploid in Orychophragmus violaceus

Orychophragmus violaceus is extremely valuable as the ornamental, vegetable, forage and oil germplasm resource. In order to develop the technique of microspore-derived embryoid induction and plant regeneration, and provide dada for the origin and evolution of the genome, the effects of the heat-shock incubation duration and the content of additional activated charcoal on embryoid yield were studied through microspore culture, and the meiotic chromosome pairing behavior of the haploid was observed by conventional squashing method in O. violaceus. The activated charcoal addition and heat shock culture were required for embryoid induction. When 4 mL microspore suspension with 1 bud per mL was incubated in a Φ6 cm petri dish at 32℃ of heat shock for three days and supplemented with 1 mg activated charcoal in each dish, the cotyledon-shaped embryoid yield and total embryoid yield were highest, which were 0.92 ± 0.18 and 1.32 ± 0.25 embryoids per bud, respectively. The germination rate of the cotyledon-shaped embryoids in 1/2MS medium was 27.73%. The natural chromosome doubling rate was 25% among the survival microspore-derived plantlets. The chromosome number of the double haploid plants and the haploid plants was 24 and 12, respectively. The meiotic chromosome pairing configuration of the haploid in O. violaceus was averaged as n = 12 = 6.352I + 2.008II + 0.384III + 0.12IV. The percentage of the pollen mother cells with bivalent, trivalent and tetravalent was up to 96%. The 12 chromosomes in 0.8% of pollen mother cells synapsed into three tetravalents. The chromosome pairing behavior strongly suggested that O. violaceus originated from a homologous octoploid with the basic chromosome number of x = 3. The above results provide a reference for breeding new materials and cultivars and for basic research in O. violaceus.