摘要
本研究以异黄酮含量不同的4个品种为试材,连续3年在5个地点种植,分析了种植年份、地点和基因型对大豆异黄酮含量的影响。用异黄酮含量不同的2个大豆品种进行了砂培试验,分析氮素和磷素对大豆异黄酮含量的影响及大豆叶片、籽粒中异黄酮含量的积累动态,同时分析了叶片中异黄酮含量变化的分子机制。在哈尔滨地区2007-2008年采用7个品种设置不同种植密度和施氮量试验,分析了农艺措施对大豆异黄酮含量的影响。研究结果如下:
1不同基因型大豆籽粒异黄酮含量生态差异研究
1.1生态条件对大豆籽粒大豆黄素含量的影响
不同种植年份对大豆黄素含量有极显著的影响。2005-2007年不同种植地点对大豆黄素含量每年都有极显著的影响,对3年结果进行联合方差分析后发现,不同种植地点对大豆黄素含量有显著的影响,高纬度有利于大豆黄素生成。2005-2007年不同基因型在每年对大豆黄素含量都有极显著的影响。互作效应对大豆黄素含量有较大影响,通过对基因型与种植地点的互作效应分析发现,不同品种对种植地点有较强的选择性。影响大豆黄素含量的各因素效应大小顺序为:年份＞基因型×年份×地点＞地点＞年份×地点。
1.2不同生态条件对大豆籽粒黄豆黄素含量的影响
不同种植年份对黄豆黄素含量有显著影响。2005-2007年,每年的不同种植地点对黄豆黄素含量都有极显著的影响,高纬度也有利于黄豆黄素生成。试验中每年不同基因型对黄豆黄素含量都有极显著的影响。互作效应对黄豆黄素含量有较大影响,通过对基因型与种植地点的互作效应分析发现,不同品种对种植地点有较强的选择性。影响黄豆黄素含量的各项因素效应大小顺序为:基因型×年份×地点＞年份＞年份×地点＞基因型。
1.3不同生态条件对大豆籽粒染料木素含量的影响
不同种植年份对染料木素含量没有显著的影响。试验中每年不同种植地点对染料木素含量都有极显著的影响,高纬度有利于染料木素生成。每年不同基因型都对染料木素含量有极显著的影响,3年结果联合方差分析中,基因型对染料木素含量有显著影响。互作效应对黄豆黄素含量有较大影响,通过对基因型与种植地点的互作效应分析发现,不同品种对种植地点有较强的选择性。影响染料木素含量的各项因素效应大小顺序为:年份×地点＞地点＞年份＞基因型×年份×地点＞基因型。
1.4不同生态条件对大豆籽粒大豆异黄酮总含量的影响
不同种植年份对大豆异黄酮总含量有极显著的影响。每年不同种植地点对大豆异黄酮总含量都有极显著的影响,对3年结果进行联合方差分析后发现,种植地点对大豆异黄酮总含量有显著影响,高纬度有利于大豆异黄酮生成。不同基因型对大豆异黄酮总含量有极显著影响。互作效应对大豆异黄酮总含量有较大影响,通过对基因型与种植地点的互作效应分析发现,不同品种对种植地点有较强的选择性。影响大豆异黄酮总含量的各项因素效应大小顺序为:年份＞地点＞年份×地点＞基因型×年份×地点＞基因型。
2.栽培措施对大豆籽粒异黄酮含量的影响
2.1盆栽条件下氮素和磷素对大豆籽粒异黄酮总含量的影响
低氮素处理条件下,大豆籽粒异黄酮总含量显著高于中氮、高氮和空白处理条件下的异黄酮总含量。不同磷素处理条件大豆籽粒异黄酮总含量的变化规律存在品种间差异。低磷处理对东农51和垦鉴27的大豆籽粒异黄酮总含量均有提高的效果。在高磷处理条件下,垦鉴27大豆籽粒异黄酮总含量与中磷处理无显著差异,且其平均值还较中磷处理低;东农51在高磷处理下异黄酮总含量显著高于中磷与空白处理。
不同年份大豆籽粒异黄酮总含量对肥料处理的反应不同。低氮处理具有促进提高大豆籽异黄酮总含量的作用。但是,不同年份低氮处理较其他处理增加幅度不同。年际间大豆籽粒异黄酮总含量对磷素反应不同,2007年低磷、中磷和高磷处理条件下大豆籽粒异黄酮总含量间无显著差异,三者均显著高于空白对照。2008年低磷处理异黄酮总含量显著高于中磷、高磷和空白对照,而中磷、高磷和空白对照的异黄酮总含量间无显著差异。
2.2大田条件下栽培措施对异黄酮含量的影响
2007年和2008年单年方差分析中种植密度、施氮量、基因型和它们互作对大豆籽粒中大豆黄素含量均有极显著的影响。两年的联合方差分析中,种植密度、基因型及各项互作项对大豆籽粒中大豆黄素含量均有极显著的影响。通过本试验认为,选用大豆品种黑农37,种植密度22万株/hm~2,纯氮施用量30 kg/hm~2均可提高大豆黄素含量。由于较强的互作效应的存在,以大豆黄素含量为目标性状进行大豆栽培生产时,应针对不同品种提出其特定的配套栽培措施。
2007年和2008年单年方差分析中种植密度、施氮量、基因型及它们互作项对大豆籽粒中黄豆黄素含量均有极显著的影响。两年的联合方差分析中,种植密度、基因型及各项互作项对大豆籽粒中黄豆黄素含量均有极显著的影响。通过本试验认为,选用大豆品种黑农48,种植密度27万株/hm~2,纯氮施用量30 kg/hm~2均可提高黄豆黄素含量。由于互作效应的存在,以黄豆黄素含量为目标性状进行大豆栽培生产时,应针对不同品种提出其特定的配套栽培措施。
2007年和2008年单年方差分析中栽培密度、施氮量、基因型及它们互作项对大豆籽粒中染料木素含量均有极显著的影响。两年的联合方差分析中,种植密度、施氮量、基因型及各项互作项对大豆籽粒中染料木素含量均有极显著的影响。选用品种东农51,22万株/hm~2种植密度,30kg/hm~2施氮量均有提高染料木素含量的效果。由于互作效应的存在,染料木素含量为目标性状进行大豆栽培生产时,应针对不同品种提出其特定的配套栽培措施。
种植密度、施氮量、基因型及各项互作项对大豆籽粒中异黄酮总含量均有极显著的影响。22万株/hm~2种植密度,30kg/hm~2施氮水平,选用东农51与黑农37均有提高异黄酮总含量的效果。由于互作效应的存在,在以大豆异黄酮总含量为目标性状进行大豆栽培生产时,应针对不同品种提出其特定的配套栽培措施。
3.大豆异黄酮积累动态规律及其分子机制分析
3.1大豆发育过程中叶片异黄酮积累动态分析
在均衡营养液处理条件下,两品种在浇营养液后较浇营养液前均有一个明显的异黄酮总含量及3种组份含量的降低,并且幅度很大。而在空白对照中,在浇营养液前后异黄酮总含量及3种组份含量没有明显的增减变化。异黄酮总含量及3种组份含量均在R1前后出现一个明显的波峰。说明进入花期前后,大豆叶片中异黄酮总含量出现一个积累高峰。
3.2苯丙氨酸解氨基因相对表达量与大豆叶片中异黄酮总含量及3种苷元组份含量间有协同增减趋势。
3.3发育中的大豆籽粒异黄酮总含量及3种苷元组份含量与籽粒发育日数呈极显著正相关。
To survey the difference of isoflavones content in soybean seeds in different environment four cultivars were planted at five locations from 2005 to 2007.Two soybean cultivars were adopted in sand culture experiment,to analysis soybean seed isoflavone content changes under different N and P applied rate,and,to analysis isoflavone content sequential changes in soybean seed and leaf,meanwhile,to analysis molecular mechanism of isoflavone content change in the leaf.7 soybean cultivars were planted under different seeding rate and N-application rate in the Harbin area in 2007-2008 years,to analysis the agronomy measure influencing soybean isoflavone content.The results were as follows:
1 Ecological difference of isoflavones content in soybean seeds among different genotypes
1.1 The difference of daidzein content in different ecological condition
There were significant difference in daidzein contents among years.There were significant difference in daidzein contents among planting sites.Daidzein contents were significantly positively correlated with latitude of the locations at the 0.01 probability levels. Different genotypes influence daidzein content at the 0.01 probability levels.Interaction of the factors influenced daidzein content greaty.Relative importance of factors affecting daidzein content were as follow:Year＞genotypex yearx location＞location＞year×location.
1.2 The difference of glycitein content in different ecological condition
There were significant difference in glycitein contents among years.There were significant difference in glycitein contents among planting locations.Glycitein contents were significantly positively correlated with latitude of the locations at the 0.01 probability levels. Different genotypes influence glycitein content at the 0.01 probability levels in every year of the three years.Interaction of the factors influenced glycitein content greaty.Relative importance of factors affecting glycitein content were as follow:Genotype×year×location＞year＞year×location＞genotype.
1.3 The difference of genistein content in different ecological condition
There were significant difference in genistein contents among years.Every year,there were significant difference in genistein contents among planting locations.Genistein contents were significantly positively correlated with latitude of the locations at the 0.01 probability levels.Different genotypes influence genistein content at the 0.01 probability levels in every year of the three years,at the 0.05 probability levels in three years.Interaction of the factors influenced genistein content greaty.Relative importance of factors affecting genistein content were as follow:Year×location＞location＞year＞genotype×year×location＞genotype.
1.4 The difference of isoflavone content in different ecological condition
There were significant difference in isoflavone contents among years.Every year,there were significant difference in isoflavone contents among planting locations at the 0.01 probability levels,at the 0.05 probability levels in three years.Isoflavone contents were significantly positively correlated with latitude of the locations at the 0.01 probability levels. Different genotypes influence isoflavone content at the 0.01 probability levels in every year and three years.Interaction of the factors influenced isoflavone content greaty.Relative importance of factors affecting isoflavone content were as follow:Year＞location＞year×location＞genotype×year×location＞genotype.
2 Effect of planting measure on isoflavone contents in soybean seeds
2.1 Effects of N and P on isoflavone contents in soybean seeds under pot culture experiment.
Soybean isoflavone content under the low N condition was significant higher than the content under high N and CK.Isoflavone content changes under different P treatment existed cultivars difference.Isoflavone content were higher under low P processes in all two cultivars. But,these two cultivars exist difference in high P processes.In different years,there were different response rules of isoflavone content to N and P processes.
2.2 Effect of planting measure on isoflavone contents in soybean seeds under field trial
Seeding rate,N-application rate,genotype and their interaction influenced daidzein content significantly in 2007 and 2008.Seeding rate,genotype and all interaction influenced daidzein content significantly in union variance analysis.Through this experiment,we fond that Heinong37 cultivar,22 ten thousand plant/hm2 seeding rate,30 kg/hm2 N-application rate all could improve daidzein content.As the interaction existing,we should adopt special cultivation measure to different cultivars to improve daidzein content in soybean seeds.
Seeding rate,N-application rate,genotype and their interaction influenced glycitein content significantly in 2007 and 2008.Seeding rate,genotype and all interaction influenced glycitein content significantly in union variance analysis.Through this experiment,we fond that Heinong 48 cultivar,27 ten thousand plant/hm2 seeding rate,30 kg/hm2 N-application rate all could improve glycitein content.As the interaction existing,we should adopt special cultivation measure to different cultivars to improve glycitein content in soybean seeds.
Seeding rate,N-application rate,genotype and their interaction influenced genistein content significantly.Through this experiment,we fond that Dongnong 51 cultivar,22 ten thousand plant/hm2 seeding rate,30 kg/hm2 N-application rate all could improve genistein content.As the interaction existing,we should adopt special cultivation measure to different cultivars to improve genistein content in soybean seeds.
Seeding rate,N-application rate,genotype and their interaction influenced isoflavone content significantly.Through this experiment,we fond that Dongnong 51 and Heinong 37 cultivar,22 ten thousand plant/hm2 seeding rate,30 kg/hm2 N-application rate all could improve isoflavone content.As the interaction existing,we should adopt special cultivation measure to different cultivars to improve isoflavone content in soybean seeds.
3 Isoflavone accumulation in soybean and it's molecular mechanism analysis
3.1 Isoflavone accumulation in soybean leafs
Soybean isoflavone content reduced greatly after pouring nourishing cream.Soybean isoflavone content appeared a peak in Rl stage.
3.2 There coordination fluctuation tendency.between alanin ammonialyase(PAL) gene relative expression quantity and isoflavone content in soybean leaf.
3.3 Growth date number was significantly related to soybean seeds isoflavone content.
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