不同化学试剂和处理方式加倍小麦单倍体植株的效果

doi:10.3864/j.issn.0578-1752.2018.05.001

摘要/Abstract

摘要： 【目的】长期以来，小麦单倍体植株染色体加倍主要采用移栽前秋水仙素溶液通气浸泡分蘖节方法，该方法存在操作复杂、污染环境等问题，而且秋水仙素毒性较强、用量较大、价格较高。本研究的目的是建立小麦单倍体植株简便、安全、高效的加倍方法，寻找可以替代秋水仙素用于小麦单倍体植株加倍的化学试剂。【方法】通过小麦品系Fielder花药培养，小麦品种科农199、新春9号和小麦品系CB037、中国春（CS）与玉米自交系郑58杂交获得小麦单倍体植株，小麦品系中国春与甘肃黑麦杂交获得小麦-黑麦双单倍体植株，利用0、5、10和20 mmol·L^-1秋水仙素溶液分别采取分蘖节加注、叶片涂抹和培养基表面添加方式对不同来源小麦单倍体植株进行加倍，并采用培养基表面添加0、30、60和120 μmol·L^-1甲基胺草磷、炔苯酰草胺和氟乐灵溶液对小麦单倍体植株及小麦-黑麦双单倍体植株加倍，比较不同加倍方法和3种除草剂的加倍效果，确定各个加倍试剂的适宜浓度。【结果】不同浓度秋水仙素溶液（0、5、10和20 mmol·L^-1）加注小麦与玉米杂交单倍体植株分蘖节部位对小麦单倍体植株不具有加倍效果，不宜在小麦单倍体植株加倍中采用。10 mmol·L^-1秋水仙素溶液涂抹拔节期小麦单倍体植株叶片的加倍率为7.7%，其他3个秋水仙素溶液浓度没有加倍成功，也不适合小麦单倍体植株加倍。培养基表面添加4个浓度秋水仙素溶液处理小麦花药培养单倍体植株的加倍率分别为26.7%、42.9%、73.3%和85.7%。表明培养基表面添加秋水仙素溶液对小麦单倍体植株的加倍效果最好，适宜浓度至少为20 mmol·L^-1。培养基表面添加0、30、60和120 μmol·L^-1炔苯酰草胺溶液处理小麦与玉米杂交单倍体植株的加倍率分别为0、0—57.1%、28.6%—75.0%和0—100%，其他2种除草剂处理小麦与玉米杂交单倍体植株没有成功；培养基表面添加120 μmol·L^-1炔苯酰草胺溶液处理小麦与黑麦杂交双单倍体植株的加倍率为9.0%，添加其他3个浓度炔苯酰草胺溶液和其他3种加倍试剂均没有结实。【结论】60—120 μmol·L^-1浓度炔苯酰草胺溶液对小麦单倍体植株加倍具有较好效果，培养基表面添加适宜浓度秋水仙素和炔苯酰草胺溶液对小麦单倍体植株加倍有效，而且简单易行。

关键词: 小麦, 单倍体植株, 远缘杂交, 花药培养, 加倍试剂

Abstract: 【Objective】 The treatment of tillering node with colchicine in a pump solution has long been widely used for chromosome doubling of haploid wheat plants before transplanting. However, this method has some disadvantages, for example, complicated manipulation and environmental or container pollution. Moreover, colchicine is some toxic, and large amount of application is not cost effective. The purposes of this study were to develop a simple, safe and efficient method for chromosome doubling of wheat haploid plants, and to evaluate a chemical that is suitable for the replacement of colchicine for chromosome doubling in wheat. 【Method】 The haploid wheat plants were obtained by anther culture for wheat line Fielder, and by crossing with the maize inbred line Zheng58 for wheat cultivars/lines Kenong199, Xinchun9, Chinese Spring and CB037 via chromosome elimination. Wheat line Chinese Spring was also crossed with rye line Gansu Heimai to produce wheat-rye double haploid plants. Chromosome doubling of different wheat haploid plants was conducted using the colchicine solutions at the concentrations of 0, 5, 10, and 20 mmol·L^-1 through tiller node adding after transplanting, leaf painting after transplanting, and medium surface adding before transplanting. Additionally, the supplement of amiprophos-methyl, trifluralin, and propyzamide at the concentrations of 0, 30, 60 and 120 μmol·L^-1 on medium surface adding was used to double chromosomes of the wheat haploid plants and the wheat-rye double haploid plants. The efficiency of chromosome doubling by different treatment methods was compared and the optimal concentrations of different chemicals were determined. 【Result】 Results indicated that the application of different concentrations of colchicine (0, 5, 10, and 20 mmol·L^-1) by tiller node adding couldn’t be able to double the wheat haploid plants from the cross of wheat and maize, so it is not useful in chromosome doubling of wheat haploid plants. A doubling frequency of 7.7% was found when 10 mmol·L^-1 colchicine was applied by leaf painting for the wheat haploid plants from maize pollen induction at jointing stage, but no haploid plant was doubled with other three concentrations of colchicine solutions, suggesting that this treatment method was also not useful for the doubling of the wheat haploid plants. The frequencies of chromosome doubling for the wheat haploid plants generated from anther culture treated with the four colchicine concentrations by medium surface adding were 26.7%, 42.9%, 73.3%, and 85.7%, respectively. It is demonstrated that the supplement of colchicine solution on medium surface adding resulted in the highest efficiency of chromosome doubling for the wheat haploid plants and the optimal concentration is at least 20 mmol·L^-1. The frequency of chromosome doubling for the haploid plants generated from the cross of wheat and maize by medium surface adding of propyzamide at the concentrations of 0, 30, 60, and 20 μmol·L^-1 was 0, 0-57.1%, 28.6%-75.0%, and 0-100%. However, the other two herbicides used by this method were not effective in chromosome doubling. The frequency of chromosome doubling by medium surface adding of 120 μmol·L^-1 propyzamide for the wheat-rye double haploid plants was 9.0%. No seed was produced by the other three concentrations of propyzamide and the other three agents (amiprophos-methyl, trifluralin, and colchicine). 【Conclusion】 It is concluded that propyzamide at the concentration of 60-120 μmol·L^-1 produced promising results of chromosome doubling for the wheat haploid plants. The medium surface adding of colchicine and propyzamide at appropriate concentrations is effective and easily adopted to double chromosomes of wheat haploid plants.

Key words: wheat, haploid plants, distant hybridization, anther culture, doubling agents

尹米琦，张双喜，范春捆，王坤杨，王静，王轲，杜丽璞，叶兴国. 不同化学试剂和处理方式加倍小麦单倍体植株的效果[J]. 中国农业科学, 2018, 51(5): 811-820.

YIN MiQi, ZHANG ShuangXi, FAN ChunKun, WANG KunYang, WANG Jing, WANG Ke, DU LiPu, YE XingGuo. Effects of Different Chemicals and Treatment Methods on Chromosome Doubling of Haploid Wheat Plants[J]. Scientia Agricultura Sinica, 2018, 51(5): 811-820.

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