利用分子标记辅助选择技术创制耐铝玉米种质

Breeding aluminum-tolerant maize germplasm using molecular marker-assisted selection

  • 摘要: 【目的】将耐铝主效基因导入普通玉米自交系,筛选出耐铝玉米株系,为拓宽玉米耐铝种质资源及耐铝育种提供理论参考。【方法】以玉米耐铝材料(CML530、CML532、CML533和CML534)和普通玉米自交系(西1、9058和LX9801)为材料,构建6个回交群体,基于分子标记辅助选择技术,利用与耐铝基因ZmM1ZmM2ZmASLZmALMT2紧密连锁的分子标记(umc1468、ZmMATE1、ZmMATE2、MateF2和ALMT93496)进行靶基因选择,将耐铝主效基因导入到普通玉米自交系,结合植株田间表现,筛选出耐铝的稳定株系,并对其进行耐铝鉴定。【结果】在分离世代(BC1F1/S1)苗期对6个回交群体共581个株系进行靶基因选择,结果有69个株系中选,结合植株田间表现,最终获得17个稳定株系。基于分子标记检测结果,17个稳定株系的遗传背景恢复率为73.9%~93.8%,平均为83.4%。0.2 mmol/L AlCl3溶液处理下17个稳定株系的根系生长均受到不同程度的抑制,根净增长度平均值为0.81 cm,极显著低于对照(0.5 mmol/L CaCl2溶液)(P<0.01,下同);稳定株系的根相对伸长率与轮回亲本差异达极显著水平;17个稳定株系的根相对伸长率为64.50%~83.33%,平均为74.74%,均为中等及以上耐铝株系,其中NS1、NS9、NS10、NS15和NS16为高耐铝株系。【结论】通过分子标记辅助选择技术可将耐铝主效基因导入普通玉米自交系,且导入多个耐铝基因株系的耐铝性较同一群体中导入单个耐铝基因的株系均有不同程度提高,表明多个耐铝基因累加可提高耐铝性。可见,该技术可作为耐铝玉米种质创制的有效手段。

     

    Abstract: 【Objective】The main-effect genes for aluminum(Al) tolerance were introgressed into common maize inbred lines, maize germplasms with Al tolerance were selected so as to provide reference for broadening Al-tolerant maize germplasms and breeding of Al-tolerant maize germplasms.【Method】Six backcross populations were constructed with Altolerant maize materials(CML530, CML532, CML533 and CML534) and common maize inbred lines(Xi1, 9058 and LX9801). Based on molecular marker-assisted selection(MAS), the targeted genes were selected using molecular markers(umc1468, ZmMATE1, ZmMATE2, MateF2 and ALMT93496) closely linked to Al-tolerant genes(ZmM1, ZmM2, ZmASL and ZmALMT2). The main-effect genes for Al tolerance were introgressed into common maize inbred lines, combined with the field performance, the stable lines were selected and Al-resistance was identified.【Result】In separated generation(BC1F1/S1), 581 individuals were screened out from 6 populations at seedling stage using MAS, and a total of 69 individuals were selected. Combined with the field performance, 17 stable lines were finally obtained. The recovery rate of genetic background based on molecular markers was 73.9%-93.8%, with an average of 83.4%. Under the treatment of 0.2 mmol/L AlCl3 solution, the growth of the roots of 17 stable lines was inhibited to varying degrees. The average net growth of the roots was 0.81 cm, which was extremely significantly lower(P<0.01, the same below) than that of the control(0.5 mmol/L CaCl2 solution). The relative elongation of the roots of the stable lines showed extremely significant difference from that of the recurrent parent. The identification of Al tolerance at seedling stage showed that the relative root elongation rate of 17 selected lines ranged from 64.50%to 83.33%, with an average of 74.74%. All of them were medium or above Al-tolerant lines, NS1, NS9, NS10, NS14 and NS16 of which were highly Al-tolerant lines.【Conclusion】Molecular marker-assisted selection technique helps introgress the Al-tolerant major-effect genes into the normal maize inbred line. Moreover, the Al tolerance of inbred lines taking in more than one Al-tolerant genes is better than lines in the same population taking in a single Al-tolerant gene. Therefore, the accumulation of multiple Al-tolerant genes enhances the level of Al tolerance. The technology can serve as an effective method for the breeding of Al-tolerant maize germplasm.

     

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