【作者】 王志强；

【导师】 吕德彬；

【作者基本信息】 河南农业大学 ， 作物遗传育种， 2004， 硕士

【摘要】 本试验以豫农9676，豫农9901，豫麦18号，大粒1号，豫麦49，国麦1号为材料，分别研究了不同长度幼穗对出愈率的差异及其在继代过程中过氧化物酶和酸性磷酸酯酶酶谱的变化。结果表明：1）在以幼穗为外植体进行愈伤组织诱导中，不同基因型幼穗的半愈期之间有显著的差异，较短的半愈期形成的愈伤组织颗粒生长旺盛，增殖的较快；较长半愈期的品种形成的愈伤组织颗粒生长较缓慢，产生的愈伤组织大多呈现白色、透明、松软状。2）不同长度的幼穗出愈率有显著的影响。0.5-1.5cm长的幼穗切段最容易产生愈伤组织，增殖也快，接种后10天出愈率100%，有的颗粒直径达3mm，接种20天多数愈伤组织块达5-6mm,生长旺盛，呈现米黄色。过氧化物同工酶酶谱结果表明：0.5-1.5cm长的幼穗愈伤组织酶带比长度小于0.5cm的幼穗愈伤组织酶带多出两条酶带并且酶带颜色较深。3）Cu2+对愈伤组织的继代培养有着重要的影响。0.1mmol/L Cu2+对愈伤组织具有较好的筛选作用，能使较好的愈伤组织生长迅速，颗粒明显增加。不同基因型愈伤组织对Cu2+的忍耐程度表现出明显的差异，其中豫麦18、豫麦49对Cu2+的忍耐程度较高，保存下来的愈伤组织较多。过氧化物酶同工酶的酶谱表明： 0.1mmol/L Cu2+筛选出的愈伤组织比0.01mmol/L和1mmol/L两种浓度处理的愈伤组织明显的多出三条酶带。4）不同愈伤组织继代过程中过氧化物酶的酶谱表现出明显的差异，与幼胚相比第一代中幼穗来源的愈伤组织过氧化物酶带颜色较深，且多出了几条酶带。第四代中过氧化物酶带颜色较浅，酶带少几条酶带。不同继代次数过氧化物酶酶谱随继代次数的增加酶带减少。5）酸性磷酸酯酶酶谱的变化与过氧化物酶相似。不同继代次数酸性磷酸酯酶表现出差异，在第六代、第七代酶谱中酶带数最少。更多还原

【Abstract】 Induction rate of immature spike and isoenzyme bands variations of peroxidase and acid phophatase during induction and subculture was studied, using wheat cultivars Yunong 9901, Yumai 18, Dali 1, Yumai 49 and Guomai 1. The results showed that:1) During the introduction of immature spike, there was significant difference of half-induction period among different genotypes. The shorter in half-induction, the more flourishing in growth of the calli, vice verse. And Calli with long-induction pierced grew slowly, and the calli had an appearance of white, transparency, and softness.2) In calli induction of immature spikes, spike sized 0.5-1.5cm was induced more easily, and grew faster. 10 days after inoculation, the induction rate almost reached to 100%, and some of the calli sized 3mm in diameter. After 20 days, the size could reach to 5-6mm in diameter, and the calli showed a light yellow color. The results of phophatase isoenzyme band pattern showed that spike calli of more than 1.5cm in size had an extra band, and the band was very thick. 3) As influence factor, Cu2+ had important effect on the culture of calli. Cu2+ concentration of 0.1mmol/L had positive selective effect to the calli. Under this condition, high-quality calli grew faster and showed granule-like tissues. The endurance ability of different calli of different genotypes showed significant difference. Of the genotypes tested, Yumai 18 and Yumai 49 showed high endurance ability to Cu2+, and more calli survived. The results of peroxidase isoenzyme band pattern showed that calli selected by 0.1mmol/L Cu2+ had 2 more extra bands than the calli treated with 0.1mmol/L or 1mmol/L Cu2+.4) There was a variation of peroxidase isoenzyme band pattern among the same generation with different explants. The peroxidase isoenzyme band pattern of the first generation of immature spike was thicker than that of immature embryos, and appeared some extra bands. Of the fourth subculture generation, peroxidase enzyme band pattern of immature embryos was thicker than that of immature spike，and also showed more extra bands. Compared with other cultivars, Yumai 18 and Yumai 49 had an extra band. With increasing of subculture times, peroxidase was enzyme bands were decreasing.5) The results of acid posphorase were rather similar to that of peroxidase, and difference of acid posphorase was observed during different subcultures. And the 6th and 7th generations showed the least enzyme bands.更多还原