【作者】 温放；

【导师】 张启翔；

【作者基本信息】 北京林业大学 ， 园林植物与观赏园艺， 2008， 博士

【摘要】 苦苣苔科Gesneriaceae在全世界约有150属3 700种,我国分布有58属470余种,绝大部分种类具有较高的观赏价值。美国、加拿大等西方国家已育成大量园艺品种,然而目前我国的苦苣苔资源基本仍处于野生状态,园艺化研究较少,部分资源受各种因素影响遭到严重破坏,且国内尚未开展对苦苣苔科观赏植物种质资源的系统研究。中国从西南到华南的山地是中国苦苣苔科植物现代分布和分化的重点区域,广西正处于这个区域中心,所拥有的属数居全国之首,物种的丰富度和多样性程度都很高,对该地区进行苦苣苔科植物观赏资源的系统性研究具有十分重要的意义。针对于此,本研究于2004年开始,先后8次对广西野生苦苣苔观赏植物种质资源进行了详细的调查,初步摸清了其分布情况,在此基础上,首次将野生苦苣苔科植物观赏资源的研究从野外调查和生物学研究范畴拓展到引种栽培、生理特性和繁殖技术、观赏性状评价、亲缘关系鉴定和杂交育种等园艺方面,初步形成了一个完整的体系,为进一步的苦苣苔科观赏植物的产业化开发奠定了基础。主要研究结果如下:1.通过对广西苦苣苔科植物分布重点区域的详细调查,共调查到33个属的132种该科植物,其中74个为广西特有种;发现了唇柱苣苔属Chirita 2新种,细筒苣苔属Lagarosolen 1新种,石蝴蝶属Petrocosmea 1新种,小花苣苔属Chiritopsis 1新变种以及短筒苣苔属Boeica、石蝴蝶属、小花苣苔属和唇柱苣苔属广西新记录各1个。在充分调查和引种的基础上,通过对2个近缘属——唇柱苣苔属和小花苣苔属的综合评价以及11个属74个种的层次分析评价,首次建立了该科植物观赏资源的评价体系,认为以唇柱苣苔属、小花苣苔属、朱红苣苔属Calcareoboea、细筒苣苔属、旋蒴苣苔属Boea、长檐苣苔属Dolicholoma、石蝴蝶属、石山苣苔属Petrocodon以及芒毛苣苔属Aeschynanthus等9个属的36个种及2变种,具有很高的观赏价值和开发潜力,其中唇柱苣苔属大部分种适应能力强,花大色艳,在评价体系中名列前茅,共有27种1个变种值得优先开发。按照生境类型将野外苦苣苔科植物划分为9类,即:阴湿半附生岩生型、洞穴型、耐旱喜光型、阴生干燥型、高海拔型、低海拔高温干燥型、阴湿喜阴附生型、耐旱附生型和林下地生型,这对成功引种该科植物具有重要的指导意义。在调查的过程中了解到大部分的苦苣苔科植物分布狭窄,居群数量少,自然更新能力弱,急需采取相应的保护措施。2.在详细调查的基础上,建立了一套引种驯化技术体系,其中引种成功的有唇柱苣苔属、小花苣苔属、朱红苣苔属、细筒苣苔属、吊石苣苔属Lysionotus和半蒴苣苔属Hemiboea等11个属,67个种。对引种成功的35种唇柱苣苔属植物的光合作用特性研究表明,唇柱苣苔属植物的光饱和点和补偿点按照其原始生长环境不同而区别明显,大部分唇柱苣苔属植物光补偿点、光饱和点和表观量子效率都较一般耐阴植物低,显示出对阴生环境的高度适应性,其中光补偿点最低的为洞穴植物软叶唇柱苣苔C. mollifolia,仅为0.57μmol·m-2·s-1,光饱和点最高的的线叶唇柱苣苔C. linearifolia也仅为16.73μmol·m-2·s-1,低于一般常见的耐阴园林植物;叶绿素a、b含量和比值试验结果显示,除少数几个种外,大部分的唇柱苣苔属和其他苦苣苔科植物的叶绿素a/b比值都在2.3左右,表现出了阴生植物的特点,因此在引种驯化过程中尤其要注意保持适当的遮阴。同时这些特点都使得该类植物具有了作为室内观赏盆花的极大潜力。3.以牛耳朵C. eburnea、黄花牛耳朵C. lutea、紫纹唇柱苣苔C. pseudoeburnea和桂林唇柱苣苔C. gueilienensis等为代表的唇柱苣苔属植物以及滇桂喜鹊苣苔Ornithoboea wildeana、吊石苣苔L. pauciflorus的种子在25~30℃时有较高的萌发率;浸种时间以16 h最佳;赤霉素对种子萌发影响不大;光照能促进种子的萌发,赤霉素处理能部分替代种子萌发对光的需求。以唇柱苣苔属为主要材料,叶插繁殖方式的研究结果表明具有肥厚肉质叶的莲座型种叶插繁殖成活率均可达100%,但繁殖系数低,更适于家庭繁殖使用;在组织培养中,不同种的不同外植体取材和最佳灭菌时间均不同,须按照实际情况进行试验和选择。均以采用2次0.1%Hgcl2灭菌为宜,幼叶和幼嫩花梗的灭菌时间均为3 min和5~7min;封闭性组织如苞片内和花蕾内组织,灭菌时间为3 min和5 min左右能得到较理想的结果;在外植体启动培养基的激素配比上,使用MS培养基,唇柱苣苔属植物多适合于NAA0.1mg·L-1+6-BA 0.1mg·L-1的激素配比,而小花苣苔属更适合于NAA0.1 mg·L-1+6-BA0.2 mg·L-1的激素配比。对7种唇柱苣苔和1种小花苣苔进行了试管苗出瓶移栽方法的研究,以蛭石40%,珍珠岩40%与草炭20%的混合基质配比,多菌灵800倍液处理基质,出瓶时间为春秋两季能获得最高的成活率;通过对不同配比的栽培基质与3种唇柱苣苔属植物生长因子之间的相互影响关系研究,认为对不同产地环境条件下的种其适应的基质配比不一,其中牛耳朵和尖萼唇柱苣苔C. pungentisepala适合于草炭60%,珍珠岩20%和蛭石20%的混合基质,桂林唇柱苣苔适合于草炭80%和珍珠岩20%的混合基质。在此基础上,建立了一套以唇柱苣苔属种为主体的引种驯化和温室盆栽技术体系,包括幼苗、成苗和开花期的栽培管理技术、环境控制、病虫害防治措施等,同时建成了国内搜集唇柱苣苔属植物比较齐全的种质资源圃。4.对56个唇柱苣苔属植物的材料进行了AFLP标记和聚类分析,试验表明,根据亲缘关系的远近,在遗传距离为0.56处分为2大类,在遗传距离0.585处,可将余下的30份材料分为2大类。对唇柱苣苔属牛耳朵、蚂蝗七C. fimbrisepala、永福唇柱苣苔C. yungfuensis和尖萼唇柱苣苔的新鲜花粉和不同液氮冻存时间花粉的萌发试验表明液氮冻存是该属植物有效的花粉保存方式,冻存前后花粉萌发率均可达70%以上;在对亲缘关系进行分析的基础上,对所搜集的唇柱苣苔属植物进行了初步的杂交育种,并获得了杂种F1后代,并对亲本重要观赏性状在子代上的表现和控制性状的基因的显隐性关系进行了分析。试验结果显示,即使亲缘关系较远的唇柱苣苔属内种,其种间杂交没有明显的生殖隔离的现象,这为培育新品种提供了保证。总之,通过引种栽培,对野生苦苣苔科植物,尤其是唇柱苣苔属内观赏资源的引种驯化、盆花栽培提供理论依据,为将来开发商品盆花以及产业化栽培打下基础,AFLP分析结果为种间亲缘关系的确定和杂交亲本的选择和育种提供了参考,杂交亲和性试验显示了唇柱苣苔属植物育种的巨大潜能。更多还原

【Abstract】 There are about 150 gunes, 3 700 species in Gesneriaceae in the world, of which approximately 58 genus and 470 species distribute in China, and most of them are famous ornamental perennials with beautiful flowers and leaves, and can be cultivated as pot flowers and garden plants in warm regions. A lot of perfect horticultural varieties have been relased in many western developed countries, for example American, Canada, etc. But up to now almost all Chinese ornamental precious germplasm resources are still leaved in wild without sufficient researches on the breeding and horticultural culture. Some resources were seriously destroyed because of various factors. And the same time, researches of the ornamental resources of Gesneriaceae have been developed few so far. The mountainous areas from southwest to south in China are the very important areas of modern distributions and differentiation of Gesneriaceae plants. Guangxi is lying in the centre of these areas. The genus of which it owns of Gesneriaceae is the most in China. The diversities and richness of species in Gesneriaceae are very high degree in Guangxi. It is very great significance of systematic researches for ornamental resources of Gesneriaceae in Guangxi. 8 investigations on the wild Gesneriaceae were made in Guangxi successively from 2004. Based on the wild investigations in Guangxi, the researches were extended from the characteristic of botany in Gesneriaceae to the cultivation of introductions and domestication, reproduction technologies, relationship analysis of some species in Chirita, test of cross-breeding, etc. It has basically been established a relatively system for further industrialized operation. The results are included as follows:1. Through detail investigation of important distributions of Gesneriaceae plants in Guangxi, 33 genus and 132 species were found, of these 74 endemic species. 2 new species in Chirita, 1 new species in Lagarosolen, 1 new species in Petrocosmea, 1 variety in Chiritopsis, and one each new records of Boeica, Petrocosmea, Chiritopsis and Chirita. Based on full investigations and introduction of those plants, the comprehensive ornamental characteristics of 2 relatives’genus, Chirita and Chiritopsis, were evaluated. Further, 11 genus and 74 species were valuated by AHP. The evaluation system of ornamental resources of Gesneriaceae plants was firstly established. The results show that some genus, for example Chirita, Chiritopsis, Calcareoboea, Lagarosolen, Boea, Dolicholoma, Petrocosmea, Petrocodon,and Aeschynanthus, etc, and 36 species and 2 varieties, have a high ornamental value and potential for development. The performances of most species in Chirita are so perfect in the evaluation system. There are 27 species and 1 variety of development should be priority. In the course of that investigation, the distributions of most of Gesneriaceae plants are very narrow, populations of the number are small, and natural regeneration capacities are very weak. Some urgent protective measures should adopt as early as possibly.2. Based on the full investigation of Gesneriaceae plants in Guangxi, a set of technical system was found for introduction and domestication. 67 species in 11 genus, for example Chirita, Chiritopsis, Calcareoboea, Lagarosolen, Lysionotus, and Hemiboea, etc. were successfully introduced. The research results of photosynthesis of 35 species in Chirita showed that there was a clear distinction of the light compensation point (LCP) and light saturation point (LSP) according to the original habitats of different species. Compare with general shade-tolerance plants, the Compare with general shade-tolerance plants, the LCP, LSP and apparent quantum yield (φ) are lower that them. This results show the high degree adaptability in this kinds of plants to shade-environments. The LCP of C. mollifolia is the lowest, is only 0.57μmol·m-2·s-1, and the highest LSP in 35 species, of C. linearfolia is only 16.73μmol·m-2·s-1. The results of chlorophyll a, b content and their ratio show that almost all species in Chirita and other genus are highly resistant to the shade environments, because the ratio of the chlorophyll a and b in major portion of species in tested plants is about 2.3 besides few species. So, appropriate over-shade is very important in the procedure of introduction and domestication. At the same time this characteristic shows that the great potential is well qualified as development of indoor potted.3. The results of biological and botany physiology characters research showed that, genus Chirita for the C. eburnea, C. lutea, C. pseudoeburnea and C. gueilienensis ect, as well as Ornithoboea wildeanathe, best of soaking time of those species seeds is 16 hours, but the best of soaking time of Lysionotus pauciflorus seeds is only 8 hours. For all species, the temperature for seed germination is 25~30℃. Ranging between 0 and 200 mg·L-1, high concentration of GA is beneficial to germination for all species, and light can improve germination rate. In addition, GA can replace the effect of light during germinating. Take Chirita species as major materials, the breeding ways were studied. The results show that the survival rate of leaf cutting can reach 100% in those species with fleshy rosulate leaves. But the reproducing coefficient is low. This shortcoming makes it to be a general reproducing way only adapt to family. On study of tissue culture and regeneration, different species require to select different explants and disinfect time. The peddicel and enclosing tissue are the best suitable explant for initial culture, and are using two times of 0.1% Hgcl2 as best suitable sterilization way while the most optimal hormone rate in MS medium of sprouting to Chirita is NAA0.1 mg?L-1+6-BA0.1 mg?L-1, but the best to Chiritopsis is NAA0.1 mg?L-1+6-BA0.2 mg?L-1. Based on studying of transplantation ways for tissue culture container seedlings of 7 species in Chirita and 1 species in Chiritopsis, the results showed that the transplantations mixed medium which the proportion of peatmoss, vermiculite, perlite was 60%, 20% and 20% was the best, the best disinfect way was to use 800×Carbendazim to disinfect mix medium, and an opportune time is spring and autumn. The effect between different proportion of vermiculite, perlite, and peatmoss in mixed medium and different growth factors was showed that different species require selecting different proportion. It should select appropriate rate of substrate for perfect growth in different species. The mixed medium of peatmoss, vermiculite and perlite, which the proportion are 60%, 20% and 20% were proved suitable for C. eburnea and C. pungentisepala, which the proportion are 80%, 0% and 20%. A set of introduction, domestication and greenhouse potted technology system was founded on according to Chirita, including the propagation, environment control, prevention and cure of the plant diseases and insect pests.4. In order to establish the relationship of different species of Chirita, and to provide the evidence for the future crossbreeding in molecule, clustering of UPGMA analysis and AFLP analysis were done in 56 species of Chirita. The genetic relationship among these are some different of the materials of Chirita. The results showed that the 56 materials tested can be divided into 2 categories according to the Nei’s genetic distance 0.56, 2 categories were divided in the Nei’s genetic distance 0.585 again. The way of conservation in Liquid Nitrogen of pollen of Chirita is proved to be an excellent method based on the comparison of pollen germination experiments of fresh pollen and conserved in liquid nitrogen with different time for C. eburnea, C. fimbrisepala, C. yungfuensis and C. pungentisepala. After 16 weeks in liquid nitrogen, the germination rate of pollen of 4 Chirita species is still over 70%. Based on the analysis of relationship of different species in Chirita, some species were selected for crossbreeding, and the crossbreeds were successfully got. The results showed that it could be hybridized between different species in same genus, Chirita, and reproduced hybrid (F1) even if their relationship is distant in same genus. It will give an enough provide to cultivate more new varieties.To sum up, by cultivation of some ornamental genetic resource of Gesneriaceae from Guangxi, especially in Chirita, it offered theoretical basis on application of introduction, the domestication and commercial cultivation in the future of the wild Gesneriaceae, especially in Chirita. The test of crossbreeding and the analysis of AFLP may be favorable of confirming the relationship, selecting appropriate parents and the molecular makers assisted selections. The test of cross-compatibility displayed the huge potential ability in Chirita.更多还原