紫花苜蓿诱引蜜蜂授粉遗传机理的研究
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摘要
紫花苜蓿(Medicago sativa L．)是世界农牧业发展中重要的牧草。它是严格的异花授粉植物，其授粉主要由蜂进行。紫花苜蓿影响蜜蜂拜访数量与遗传及环境因子有关，我国的研究仍处在对授粉现象的观察，并没有深入到分子水平方面的研究。本研究以10个紫花苜蓿品种为材料，对其花托直径、花冠长度、泌蜜量及花蜜糖组成成份等花部特征、蜜蜂拜访数量和种子产量等进行了研究，同时利用了随机扩增多态性DNA技术和同工酶技术对紫花苜蓿不同品种花部特征的遗传变异进行研究。结果表明：
(1)花苜蓿的单位面积花蜜量与蜜蜂拜访数量呈极显著正相关(r=0.9299~(**))，自然蜜蜂拜访数量与种子产量呈显著正相关(r=0.8716~*)。
(2)苜蓿的花托直径能比较直观地反映出其花蜜量的多少。
(3)蜜蜂的嗅觉和视觉因素是造成紫花苜蓿各品种间的蜜蜂拜访数量差异的首要因素。其次是花朵的大小，再次是花蜜量的多少，最后是花朵的颜色。
(4)蜜蜂拜访数量是决定紫花苜蓿种子产量的重要外在因素之一。而紫花苜蓿本身特性也是决定其种子产量的主要因素。
(5)在相同的环境条件下，紫花苜蓿的自然蜜蜂拜访数量主要是由紫花苜蓿的遗传基础一基因型及其表现型决定的。
(6)同工酶作为一种特殊的蛋白质，它的产生和表达不仅受到基因调控，其作用的发挥还受到细胞内、外环境、植物阶段发育和系统发育等多种因素的影响，不可能在一个组织或品种中稳定不变的表达，但是在同期取样且保持技术一致的前提下用同工酶电泳技术可以检测出品种
间的差异。同工酶标记可以将 10个紫花首精划分为三类，L173和 Ladak
首范聚成一类，Pri。e、Algongllin禾陕北首薄聚成一类，WL323、Defa、
Sanditi、Derbv和 Sitel 甘落聚成一类。
门）MID技术分析了紫花首薄遗传多样性，10个随机引物共检测
到132条扩增片段，其中多态片段102条，占总位点的76．52％；共扩增
出了2＊3条带，平均每个引物扩增13．2条带；可以将10个紫花首范品
种划分为三类，陕北首蒲独自聚成一类，Ladak、WL323、Defa和 Algonguin
首稽聚成一类，Sanditi、Derby、Prime、Sitel和 L173首渐聚成一类。
RAPD标记聚类结果和同工酶标记聚类结果具有一定的相似性，但也存在
一些不一致的情况。
抢）应用RAPD分子标记和同工酶标记技术来揭示紫花窗营各品种
的变异对其吸引自然蜜蜂拜访数量的影响都是可行的。
Alfalfa (Medicago saliva L.) is a very important forage in the development of agriculture and stock in the world. It is a plant of allogamy and its pollination relies on honey bee mainly, honey bee visiting was influenced by its genetics and environment. For the time being, domestic research had being focused on observation of pollination, haven't reached to molecular level yet. 10 lucerne varieties were used in the study, research mainly focused on floral feature, such as receptacle diameter, corolla length, nectar volume and components of sugar, natural honey bee visitation and seed yield. Mean while, random amplified polymorphic DNA and isoenzyme technology were used to study the genetic diversity of different varieties' floral characteristics. The results showed that:
(1) Lucerne nectar production was positively correlated with honey bee visitation (r=0.9299**), honey bee visitation and seed production were positively correlated either (r=0.8716*).
(2) Lucerne nectar production was reflected directly by Receptacle diameter.
(3)Honey bee scent and vision was the most important factor that affected the honey bee visitation difference between varieties, followed by floral size, nectar volume, and floral color.
(4)Honeybee visitation was the key factor that decided the seed yield of Lucerne, followed by variety characteristics.
(5)In the same environment, honey bee visitation was decided by Lucerne genetics, ie, its gene and performance.
(6)As a kind of special protein, isoenzyme's production and express was not only controlled by gene, its function was affected by environment inside and outside the cell, plant phenology etc. it's impossible to get invariable express in same variety or tissue, but the variety difference was testable under same stage and methods using isoenzyme electrophoresis technology. 10 lucerne varieties were divided into three categories by isoenzyme marker: LI73 and Ladak; Prime, Algonguin and shanbei; WL323,Defa, Sanditi, Derby and Sitel.
(7)Genetic diversity of alfalfa was studied using RAPD. Amplification of 10 random primers detected 132 amplified fragments of which 102 bands(76.52%) were polymorphic. Amplification of these 10 primers highlighted 2-13 bands. 10 lucerne varieties were divided into three categories by RAPD marker: shanbei; Ladak, WL323, Defa and Algonguin; Sanditi, Derby, Prime, Sitel and LI73. Relationships between the 10 alfalfa revealed by RAPD markers and isoenzyme marker are similar, and there are also some exceptions.
(8) It is feasible to study different lucerne varieties' effects on honey bee visitation by RAPD marker and isoenzyme marker.
(1)花苜蓿的单位面积花蜜量与蜜蜂拜访数量呈极显著正相关(r=0.9299~(**))，自然蜜蜂拜访数量与种子产量呈显著正相关(r=0.8716~*)。
(2)苜蓿的花托直径能比较直观地反映出其花蜜量的多少。
(3)蜜蜂的嗅觉和视觉因素是造成紫花苜蓿各品种间的蜜蜂拜访数量差异的首要因素。其次是花朵的大小，再次是花蜜量的多少，最后是花朵的颜色。
(4)蜜蜂拜访数量是决定紫花苜蓿种子产量的重要外在因素之一。而紫花苜蓿本身特性也是决定其种子产量的主要因素。
(5)在相同的环境条件下，紫花苜蓿的自然蜜蜂拜访数量主要是由紫花苜蓿的遗传基础一基因型及其表现型决定的。
(6)同工酶作为一种特殊的蛋白质，它的产生和表达不仅受到基因调控，其作用的发挥还受到细胞内、外环境、植物阶段发育和系统发育等多种因素的影响，不可能在一个组织或品种中稳定不变的表达，但是在同期取样且保持技术一致的前提下用同工酶电泳技术可以检测出品种
间的差异。同工酶标记可以将 10个紫花首精划分为三类，L173和 Ladak
首范聚成一类，Pri。e、Algongllin禾陕北首薄聚成一类，WL323、Defa、
Sanditi、Derbv和 Sitel 甘落聚成一类。
门）MID技术分析了紫花首薄遗传多样性，10个随机引物共检测
到132条扩增片段，其中多态片段102条，占总位点的76．52％；共扩增
出了2＊3条带，平均每个引物扩增13．2条带；可以将10个紫花首范品
种划分为三类，陕北首蒲独自聚成一类，Ladak、WL323、Defa和 Algonguin
首稽聚成一类，Sanditi、Derby、Prime、Sitel和 L173首渐聚成一类。
RAPD标记聚类结果和同工酶标记聚类结果具有一定的相似性，但也存在
一些不一致的情况。
抢）应用RAPD分子标记和同工酶标记技术来揭示紫花窗营各品种
的变异对其吸引自然蜜蜂拜访数量的影响都是可行的。
Alfalfa (Medicago saliva L.) is a very important forage in the development of agriculture and stock in the world. It is a plant of allogamy and its pollination relies on honey bee mainly, honey bee visiting was influenced by its genetics and environment. For the time being, domestic research had being focused on observation of pollination, haven't reached to molecular level yet. 10 lucerne varieties were used in the study, research mainly focused on floral feature, such as receptacle diameter, corolla length, nectar volume and components of sugar, natural honey bee visitation and seed yield. Mean while, random amplified polymorphic DNA and isoenzyme technology were used to study the genetic diversity of different varieties' floral characteristics. The results showed that:
(1) Lucerne nectar production was positively correlated with honey bee visitation (r=0.9299**), honey bee visitation and seed production were positively correlated either (r=0.8716*).
(2) Lucerne nectar production was reflected directly by Receptacle diameter.
(3)Honey bee scent and vision was the most important factor that affected the honey bee visitation difference between varieties, followed by floral size, nectar volume, and floral color.
(4)Honeybee visitation was the key factor that decided the seed yield of Lucerne, followed by variety characteristics.
(5)In the same environment, honey bee visitation was decided by Lucerne genetics, ie, its gene and performance.
(6)As a kind of special protein, isoenzyme's production and express was not only controlled by gene, its function was affected by environment inside and outside the cell, plant phenology etc. it's impossible to get invariable express in same variety or tissue, but the variety difference was testable under same stage and methods using isoenzyme electrophoresis technology. 10 lucerne varieties were divided into three categories by isoenzyme marker: LI73 and Ladak; Prime, Algonguin and shanbei; WL323,Defa, Sanditi, Derby and Sitel.
(7)Genetic diversity of alfalfa was studied using RAPD. Amplification of 10 random primers detected 132 amplified fragments of which 102 bands(76.52%) were polymorphic. Amplification of these 10 primers highlighted 2-13 bands. 10 lucerne varieties were divided into three categories by RAPD marker: shanbei; Ladak, WL323, Defa and Algonguin; Sanditi, Derby, Prime, Sitel and LI73. Relationships between the 10 alfalfa revealed by RAPD markers and isoenzyme marker are similar, and there are also some exceptions.
(8) It is feasible to study different lucerne varieties' effects on honey bee visitation by RAPD marker and isoenzyme marker.
引文
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