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随着分子标记技术的快速发展,分子标记辅助育种已被广泛应用于玉米育种,迫切需要高通量、高效的技术方法。基于单粒种子苗前的分子生物学分析来预测表型性状是提高分子标记辅助选择效率的方法,但还需要发展一种切实有效的方法来保证苗前的分子生物学分析和种子播种后的正常发芽及生长。本研究通过对玉米单粒种子进行切取部分胚乳取样,对样品进行CTAB和碱煮法DNA提取的比较,而后对切去部分胚乳的种子进行包衣处理,然后对正常种子(T1)、只切除部分胚乳的种子(T2)、只包衣的种子(T3)以及既切除部分胚乳又包衣的种子(T4)进行室内沙盘发芽试验和田间种植试验分析,结果表明通过包衣的方法可以提高切除部分胚乳种子的发芽率,室内和田间试验中T4处理和T1处理的发芽率、株高及简易活力指数基本相近,差异均不显著。T4处理在室内和田间试验中的发芽率比T2处理分别提高了3.35%和14.90%,差异分别达到显著和极显著水平;T4处理的株高和简易活力指数均比T2处理的高,差异均达到了极显著水平。切除部分胚乳又包衣种子的发芽率高于国家标准发芽率,十天后的株高、简易活力指数基本达到正常种子的水平。对切除部分胚乳的种子包衣的方法可以解决播种前的分子生物学方面的检测又能使种子正常的发芽及生长,大大提高了玉米分子育种效率。
With the rapid development of molecular marker technology, molecular marker-assisted breeding has been widely used in maize breeding, and there is an urgent need for high-throughput and efficient technical methods. Predicting phenotypic traits based on single-seed pre-emergence molecular biology analysis is a way to improve the efficiency of molecular marker-assisted selection. However, there is still a need to develop a practical and effective method to ensure pre-emergence molecular biology analysis and post- Normal germination and growth. In this study, some endosperm samples were cut from single seeds of maize and the samples were compared with DNA extracted by CTAB and alkali-boiled method. Then the seeds of partial endosperm were coated, and then the normal seeds (T1) Partial endosperm seed (T2), coating only seed (T3) and both excised part of endosperm and seed (T4) were subjected to indoor sand table germination test and field planting test analysis. The results showed that the method of coating can improve the excision The germination percentage of some endosperm seeds, the germination rate, plant height and simple vigor index of T4 and T1 treatments in laboratory and in field were similar, but the differences were not significant. Compared with T2 treatment, the germination rate of T4 treatment increased by 3.35% and 14.90% respectively in indoor and field experiments, the difference reached significant and extremely significant level respectively. The T4 plant height and simple vigor index were higher than those of T2 treatment Reached a very significant level. Excised part of the endosperm and seed coating germination rate is higher than the national standard germination rate, ten days after the plant height, simple vitality index basically reached the level of normal seeds. The method of removing the seed coat of endosperm can solve the problem of molecular biology before sowing and normal germination and growth of the seed and greatly improve maize molecular breeding efficiency.