云南引种辣木繁育系统的研究
吴疆翀
学位类型博士
导师顾志建 ; 龚洵
2010-05
学位授予单位中国科学院研究生院
学位专业植物学
摘要辣木(Moring oleifera Lam)是辣木科(Moingaceae)辣木属(Moringa)的一种原产印度的多用途树种,具有很高的经济价值。其叶片含有丰富的蛋白质、维生素和矿物质,可用作蔬菜或饲料;种子粉含絮凝蛋白,可用作净水剂;种子油可供食用、化妆品用或作润滑油。目前辣木已广泛引种于世界各地,加之其优良的速生性状,被认为是一种重要的混农林间作树种。掌握植物的生殖生物学特性是良种培育的基础,全面和深入地了解辣木的繁育系统各组成部分可以为育种技术措施的制定提供科学依据。本文采用传统实验手段与现代分子标记相结合,对云南省引种栽培的辣木种群的花发育、开花式样、花期物候、访花昆虫种类、访花行为、访花频率进行了观察;运用石蜡切片、荧光染色、杂交指数、花粉-胚珠比、SSR分子标记方法,对辣木的繁育系统各组成部分进行了研究。研究结果如下:1. 花芽发育可分为苞片分化期、萼片分化期、花瓣分化期、雄蕊分化期和雌蕊分化期。花药发育为基本型,小孢子减数分裂为同时型。胚囊发育为蓼型。辣木雌雄生殖结构发生异常的比例很低,不阻碍生殖成功。2. 个别植株全年开花,种群每年春季和秋季开花两次,各花期分别持续约两个月,其中盛花期分别持续约一个月。单花花期持续7天,花集中在上午开放。花药先熟,TTC法测定的花粉活性持续时间为花开放至开花后48h;联苯胺-过氧化氢法测定的柱头可授期为开花后24h-72h。成熟的花药和柱头在空间和时间上分离。花的香味在开花后即出现,并可持续48h。3. 杂交指数为5、花粉-胚珠比为988.9±564.4,繁育系统类型为异交为主,部分自交亲和。自交不亲和性体现为配子体自交不亲和。4. 利用FIASCO法开发了20对具多态性的辣木SSR引物。各位点的等位基因数为2-6,平均为3。期望杂合度(HE)为0.3608至0.7606,平均为0.5455。表观杂合度(HO)为0.0000至0.8750,平均为0.4562。20个位点中有7个显著偏离哈迪-温伯格平衡。5. 利用SSR分子标记进行父本分析检测异交率和基因流。选取8对引物对试验种群12株母本和60株父本进行父本分析,在95%严格置信度条件下,可以为总共288粒种子中的155粒确定父本。多位点异交率tm=0.797,单位点异交率ts=0.742。传粉集中在以母本为圆心,半径20m的范围内。来自主风方向和逆风方向的传粉在数量上无差异。6. 生产性栽培条件下结实率较低,结实情况受传粉者限制,最主要的传粉昆虫是紫木蜂(Xylocopa valga)和黄叶带土蜂(Scolia vittifornis)。7. 生产上可以通过人工辅助异株授粉提高结实量。
资助项目Moringa oleifera Lam. (Moringaceae) is an economically important multi-purpose tree indigenous to northwest India. Featured by richness in proteins, minerals and Vitamins, leaves of M. oleifera are used as highly nutrient vegetable and cattle fodder. Besides, the seed powder is used in water purification, and the seed oil is acquired for edibles, lubricating and cosmetics. Due to its multiple applications and commercial benefits, M. oleifera has been broadly introduced and cultivated around the world, and has been identified as the important one in agri-horti-silviculture programs. Mastering the reproductive characteristics and bionomics of a species is the foundation of fine variety breeding. And understanding the breeding system of M. oleifera provides basic evidence for the establishment of breeding techniques. Both traditional methods and modern DNA marker were applied to study the component parts of breeding system of M. oleifera introduced to Yunnan, China. Floral development, anthesis phenology, flowering pattern, species and visiting frequency of pollinating insects, as well as foraging behavior of pollinators were observed. Furthermore, the type of breeding system, outcrossing rate and gene flow were also tested by means of fluorescence, paraffin sections, outcrossing index, pollen-ovule ratio, and microsatellites. Then the findings are as follows. 1. Morphological differentiation of flower bud could be divided into 5 stages: bract differentiation, sepal differentiation, petal differentiation, stamen differentiation, and pistil differentiation. Abnormality of male and female reproductive structure is rare that do not prevent successful breeding. 2. With a few individuals flowering throughout the year, florescence of population appears twice a year, respectively in spring and autumn. Each blooming period lasts about 2 months, among which the stage of full blossom lasts about 1 month. The blooming period of a single flower is 7d, and anthesis time is forenoon. Pollen viability lasts from blooming to 24h after flowering, tested by TTC. While stigma reception lasts from 24h to 72h after blooming, tested by benzidine and hydrogen peroxide. Mature anthers and stigma are apart from time and space. Flavor rises up right away after blossom and continues to 48h.3. The OCI is 5, and P/O is 988.9±564.4. The breeding system of M. oleifera is outcrossing, partially self-compatible, and demand for pollinators. Self-incompatibility is gametophytic.4. A total of twenty polymorphic microsatellite markers were developed by method of FIASCO. The number of alleles per locus ranged from two to six, with an average of three. The expected (HE) and observed (HO) heterozygosities ranged from 0.3608 to 0.7606 (average of 0.5455) and from 0.0000 to 0.8750 (average of 0.4562), respectively. Seven loci were significantly deviated from Hardy-Weinberg equilibrium.5. Paternity analysis by SSR was used to estimate the outcrossing rate and gene flow of M. oleifera. The analysis was carried out in an experimental population with 12 maternal trees and 60 paternal trees. 155 seeds out of 288 seeds were confirmed pollen donors by 8 microsatellite loci at 95% strict confidence level. The multilocus outcrossing rate is tm=0.797,and single-locus outcrossing rate is ts=0.742. Most of pollen dispersal is within 20m, and the amount of downwind distribution is not significantly distinct from the upwind.6. The natural fruiting rate of M. oleifera is low under scale cultivation, and is limited by pollinators. Most reliable pollinators are Xylocopa valga andScolia vittifornis.7. Artificial xenogamy could improve fruit setting and the yield of seeds in practice.
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/16050
专题昆明植物所硕博研究生毕业学位论文
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吴疆翀. 云南引种辣木繁育系统的研究[D]. 中国科学院研究生院,2010.
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