仁用杏新品种‘围选1号’授粉特性研究
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摘要
在仁用杏实际生产中,由于春季开花早,极易遭受晚霜的危害。而新品种‘围选1号’(Prunus armeniaca‘Weixuan 1’)是从抗寒品种中选育出的优株,抗寒性优于当地的主栽品种。本研究以‘围选1号’为母本,分别采用不同生态区的杏品种作为父本,对‘围选1号’进行授粉,通过测定其花粉生活力、柱头可授性及花粉管生长情况,不同授粉组合对坐果率、结实率及果仁品质的影响,并运用蛋白质组学方法分别对‘围选1号’和‘龙王帽’自花授粉花柱蛋白质进行比较,主要研究结果如下:
1.通过对‘围选1号’自花和异花授粉试验研究,结果表明:‘围选1号’自花结实率为2.17%(<6%),属于自交不亲和品种。在异花授粉试验中,以‘十片红’对‘围选1号’进行授粉效果最好,结实率最高,为74.88%;在授粉试验中以花束状果枝的结实率最高,长果枝最低。
2.采用过氧化物酶染色法对不同授粉品种花粉活力进行测定,结果表明:花粉生活力都在50%以上,‘围选1号’的花粉生活力高达77.81%,但花粉萌发率相对较低。
3.从联苯胺-过氧化氢法检验‘围选1号’柱头的可授性,可以看出柱头可授性在开花3 h后可授性开始增强,且能持续3 d。从人工授粉田间试验结果表明:开花当天,柱头可授性最高为30.67%,柱头可授性的强弱与其颜色之间的关系为:柱头为黄色时,可授性强。随着可授性的减弱,柱头变成黄褐色,当柱头变为深褐色时失去可授性。
4.采用荧光显微镜压片法观察不同授粉组合4 h、8 h、24 h、48 h、60 h、72 h后花柱内花粉管生长情况,发现‘围选1号’自花授粉花粉管生长受到抑制的部位大约是花柱中下部位。不同授粉组合花粉管生长初期相同,并无明显差异。但是当授粉48 h后花粉管到达柱头距子房约1/2位置处时,‘围选1号’自花授粉的花粉管末端膨大,只生长至花柱近中部就停止生长,花粉管生长受到抑制,不能进入子房,完成受精作用。而异花授粉可以继续生长,在授粉72 h后能进入子房,完成受精。
5.酚-甲醇/醋酸铵沉淀法得到的蛋白点比TCA/丙酮沉淀法所获得的蛋白点更清晰易辨、圆润、分布均匀,适合后续分析。选择‘龙王帽’自花授粉与‘围选1号’自花授粉后雌蕊中表达丰度较高且差异明显的12个蛋白质点,通过MALDI-TOF-TOF/MS对所获得的肽段进行分析,有8个蛋白得到了有效的鉴定,其中包括‘围选1号’自花授粉当中与植物自交不亲和密切相关的S-locus F-box蛋白与S-RNase酶。
6.通过不同杏品种对‘围选1号’进行人工授粉,并对其授粉后代果实出仁率、出核率、外观、重量、营养成分等方面进行初步比较。结果显示:不同组合授粉后代果实出核率、出仁率、单仁重明显高于对照‘围选1号’自然授粉,并以鲜食杏对‘围选1号’授粉后代表现较为优异,以‘围选1号’ב木瓜杏’授粉组合的出核率最高,为15.89%。‘围选1号’ב凯特’授粉组合的出仁率最高,为45.17%,不同授粉组合与对照营养成分之间变化规律不明显。
In the production of the kernel apricot, Due to the early spring blossom, it is vulnerable for it to get frost damage. New varieties Prunus armeniaca‘Weixuan 1’belongs to the hybrid offspring of the Chinese common apricots, which we selected from the cold hardiness varieties. Through grafting propagation, it has a stronger ability of resisting late frosts than local varieties. In this paper, pollination experiments and the effects of pollination tree on fruiting rate of Prunus armeniaca were studied by choosing grafting‘Weixuan 1’as female parent and different apricot tree as male parent. In order to know and master the best pollination period and pollination trees, we tested the pollen viability, stigma receptivity and the growth of pollen tube, and different pollination trees on influence of fruiting rate, seed setting rate and fruit quality. Besides, with proteomic methods, contained a comparative study on style protein between self-pollination‘Weixuan
1’and self-pollination‘Longwang mao’. The specific results is drown as follows: 1. Studying self-pollination and cross pollination composes in‘Weixuan 1’. The result is drown as follows: the fruit setting rate is respectively 2.17%(<6%), which belongs to the self-incompatible variety. As the best pollination tree of‘Weixuan 1’, fruit setting rate is‘Shipianhong’(74.88%), and the highest seed rate of flower bunch like branches, the lowest is long fruit branch.
2. Pollen viability of different varieties by pollen germination test in vitro. The result is drown as follows: varietal pollen viability is more than 50%,‘Weixuan 1’pollen viability high amounts to 84.31%, but germination ability of the pollen is relatively low.
3. It is feasibility to test stigma receptivity by the enzidine-H2O2 also. Stigma receptivity of‘wei xuan1’start to increase after 3 h blossom, which could last for 3 d. On the first day, stigma receptivity came to a maximum of 30.67%, The relationship between stigma receptivity and color is: the receptivity is strong when stigma is yellow. As the ability weakened, stigma become brown, and pollination can’t be done while the stigma is dark brown.
4. Using fluorescence spectrum technology observation, After pollinated 4 h, 8 h, 24 h, 48 h, 60 h, 72 h, pollen tubes traveling along the style in Katy could grow into ovary, and most of them in‘Weixuan 1’could stop growing with tip expanded into ball. I discovered that the pollen of self pollinated varieties on stigmas are same with the pollen of self-pollination and cross pollination composes in the pollen germination and pollen tube growth initial stage. But the pollen tube reached to the column from the ovary ca. 3/4 position 48 h after pollination, and the pollen of self pollination tube inflated in the end, stopped growing, and only a very few can reach to ovary normally. While the cross pollination could continue to grow, can enter the ovary 72 h after pollination.
5. By two-dimensional electrophoresis separation, the Protein spots we got by phenol and methanol /ammonium acetate precipitation method were more suitable for the following analysis than that by TCA / acetone precipitation method, moreover, the spots could be separated better: clear, smooth, uniform distribution. Through the MALDI-TOF- TOF/MS analysis of different protein spots. Among these 12 proteins, 8 proteins were identificated effectively. Such as proteins related to plant self-incompatibility metabolism of S-RNA enzymes (11, S-the RNase) and self-incompatibility protein (8, S-locus F-box protein 4)etc.
6. Through compared to different artificial pollination combinations, and the pollination offspring fruit yield, appearance, weight, nutritional etc. The results is drown as follows: Pollination progenies from the nucleation rate, the rate of out-jen, kernel of apricot were significantly higher than that of control‘Weixuan 1’natural pollination, Fruit shape index is moderate,‘Weixuan 1’בMuguaxing’pollination combination of nuclear rate is highest(15.89%).‘Weixuan 1’בKaty’pollination combination kernel ratio is highest(45.17%), After pollination, there was no significant difference in nutritional components of different varieties.
1.通过对‘围选1号’自花和异花授粉试验研究,结果表明:‘围选1号’自花结实率为2.17%(<6%),属于自交不亲和品种。在异花授粉试验中,以‘十片红’对‘围选1号’进行授粉效果最好,结实率最高,为74.88%;在授粉试验中以花束状果枝的结实率最高,长果枝最低。
2.采用过氧化物酶染色法对不同授粉品种花粉活力进行测定,结果表明:花粉生活力都在50%以上,‘围选1号’的花粉生活力高达77.81%,但花粉萌发率相对较低。
3.从联苯胺-过氧化氢法检验‘围选1号’柱头的可授性,可以看出柱头可授性在开花3 h后可授性开始增强,且能持续3 d。从人工授粉田间试验结果表明:开花当天,柱头可授性最高为30.67%,柱头可授性的强弱与其颜色之间的关系为:柱头为黄色时,可授性强。随着可授性的减弱,柱头变成黄褐色,当柱头变为深褐色时失去可授性。
4.采用荧光显微镜压片法观察不同授粉组合4 h、8 h、24 h、48 h、60 h、72 h后花柱内花粉管生长情况,发现‘围选1号’自花授粉花粉管生长受到抑制的部位大约是花柱中下部位。不同授粉组合花粉管生长初期相同,并无明显差异。但是当授粉48 h后花粉管到达柱头距子房约1/2位置处时,‘围选1号’自花授粉的花粉管末端膨大,只生长至花柱近中部就停止生长,花粉管生长受到抑制,不能进入子房,完成受精作用。而异花授粉可以继续生长,在授粉72 h后能进入子房,完成受精。
5.酚-甲醇/醋酸铵沉淀法得到的蛋白点比TCA/丙酮沉淀法所获得的蛋白点更清晰易辨、圆润、分布均匀,适合后续分析。选择‘龙王帽’自花授粉与‘围选1号’自花授粉后雌蕊中表达丰度较高且差异明显的12个蛋白质点,通过MALDI-TOF-TOF/MS对所获得的肽段进行分析,有8个蛋白得到了有效的鉴定,其中包括‘围选1号’自花授粉当中与植物自交不亲和密切相关的S-locus F-box蛋白与S-RNase酶。
6.通过不同杏品种对‘围选1号’进行人工授粉,并对其授粉后代果实出仁率、出核率、外观、重量、营养成分等方面进行初步比较。结果显示:不同组合授粉后代果实出核率、出仁率、单仁重明显高于对照‘围选1号’自然授粉,并以鲜食杏对‘围选1号’授粉后代表现较为优异,以‘围选1号’ב木瓜杏’授粉组合的出核率最高,为15.89%。‘围选1号’ב凯特’授粉组合的出仁率最高,为45.17%,不同授粉组合与对照营养成分之间变化规律不明显。
In the production of the kernel apricot, Due to the early spring blossom, it is vulnerable for it to get frost damage. New varieties Prunus armeniaca‘Weixuan 1’belongs to the hybrid offspring of the Chinese common apricots, which we selected from the cold hardiness varieties. Through grafting propagation, it has a stronger ability of resisting late frosts than local varieties. In this paper, pollination experiments and the effects of pollination tree on fruiting rate of Prunus armeniaca were studied by choosing grafting‘Weixuan 1’as female parent and different apricot tree as male parent. In order to know and master the best pollination period and pollination trees, we tested the pollen viability, stigma receptivity and the growth of pollen tube, and different pollination trees on influence of fruiting rate, seed setting rate and fruit quality. Besides, with proteomic methods, contained a comparative study on style protein between self-pollination‘Weixuan
1’and self-pollination‘Longwang mao’. The specific results is drown as follows: 1. Studying self-pollination and cross pollination composes in‘Weixuan 1’. The result is drown as follows: the fruit setting rate is respectively 2.17%(<6%), which belongs to the self-incompatible variety. As the best pollination tree of‘Weixuan 1’, fruit setting rate is‘Shipianhong’(74.88%), and the highest seed rate of flower bunch like branches, the lowest is long fruit branch.
2. Pollen viability of different varieties by pollen germination test in vitro. The result is drown as follows: varietal pollen viability is more than 50%,‘Weixuan 1’pollen viability high amounts to 84.31%, but germination ability of the pollen is relatively low.
3. It is feasibility to test stigma receptivity by the enzidine-H2O2 also. Stigma receptivity of‘wei xuan1’start to increase after 3 h blossom, which could last for 3 d. On the first day, stigma receptivity came to a maximum of 30.67%, The relationship between stigma receptivity and color is: the receptivity is strong when stigma is yellow. As the ability weakened, stigma become brown, and pollination can’t be done while the stigma is dark brown.
4. Using fluorescence spectrum technology observation, After pollinated 4 h, 8 h, 24 h, 48 h, 60 h, 72 h, pollen tubes traveling along the style in Katy could grow into ovary, and most of them in‘Weixuan 1’could stop growing with tip expanded into ball. I discovered that the pollen of self pollinated varieties on stigmas are same with the pollen of self-pollination and cross pollination composes in the pollen germination and pollen tube growth initial stage. But the pollen tube reached to the column from the ovary ca. 3/4 position 48 h after pollination, and the pollen of self pollination tube inflated in the end, stopped growing, and only a very few can reach to ovary normally. While the cross pollination could continue to grow, can enter the ovary 72 h after pollination.
5. By two-dimensional electrophoresis separation, the Protein spots we got by phenol and methanol /ammonium acetate precipitation method were more suitable for the following analysis than that by TCA / acetone precipitation method, moreover, the spots could be separated better: clear, smooth, uniform distribution. Through the MALDI-TOF- TOF/MS analysis of different protein spots. Among these 12 proteins, 8 proteins were identificated effectively. Such as proteins related to plant self-incompatibility metabolism of S-RNA enzymes (11, S-the RNase) and self-incompatibility protein (8, S-locus F-box protein 4)etc.
6. Through compared to different artificial pollination combinations, and the pollination offspring fruit yield, appearance, weight, nutritional etc. The results is drown as follows: Pollination progenies from the nucleation rate, the rate of out-jen, kernel of apricot were significantly higher than that of control‘Weixuan 1’natural pollination, Fruit shape index is moderate,‘Weixuan 1’בMuguaxing’pollination combination of nuclear rate is highest(15.89%).‘Weixuan 1’בKaty’pollination combination kernel ratio is highest(45.17%), After pollination, there was no significant difference in nutritional components of different varieties.
引文
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