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安徽产石蒜属植物光合特性、细胞学及总生物碱含量的研究

Studies on Photosynthetic Characteristics, Cytology and Content of Total Alkaloids of the Genus Lycoris from Anhui Province

【作者】 刘坤

【导师】 周守标;

【作者基本信息】 安徽师范大学 , 植物学, 2010, 硕士

【摘要】 石蒜属(Lycoris Herb.)是石蒜科(Amaryllidaceae)一类具鳞茎的多年生草本植物,为东亚特有属,主要分布于中国、日本和韩国。在形态解剖、细胞学、组培和繁殖、药用、遗传多样性和分子系统学等方面已经对石蒜属植物进行了大量研究,但有关石蒜属植物的光合特性的研究少有报道,对石蒜属植物药用成分的研究也主要集中在石蒜(L. radiata)等少数种上。此外,石蒜属植物的核型进化关系仍存在很大的争议。以安徽产7种石蒜属植物以及不同居群植物为研究材料,通过光合特性、核型和总生物碱含量的研究和分析,揭示了石蒜属种间及种内不同居群之间的差异,为深入探讨石蒜属植物的演化关系和核型进化趋势,以及资源的有效利用提供理论依据和基础资料。结果如下:1.研究了安徽产4种秋季出叶和3种春季出叶石蒜属植物的气体交换和叶绿素荧光参数日变化特征,发现石蒜属植物种间光合特性存在较大差异。除忽地笑(L. aurea)净光合速率PN的日变化为“单峰”型外,其余所有植物PN的日变化均为“双峰”曲线;所有植物光饱和点在322~1189μmol m-2 s-1之间,光补偿点在8.11~15.36μmol m-2 s-1之间,具有较高的表观量子效率(0.0457~0.0798)。光合特性参数表明了7种石蒜属植物均具有阴生植物的特征。2.研究了不同遮荫条件对石蒜(L. radiata var. radiata)(3n=33)生长及光合特性的影响,发现全光照下石蒜净光合速率PN的日变化为典型的“双峰”曲线,有明显的光合“午休”现象,而遮荫处理下石蒜PN日变化均为“单峰”型;60~70%的自然光下,石蒜具有相对较大的日碳固定总量、叶面积及单株总叶面积。表明了适度遮荫有利于石蒜的生长,强光和弱光对石蒜的生长均不利。3.石蒜属植物的染色体核型变异较大,研究发现了许多新的核型和染色体数目。中国石蒜(L. chinensis)在不同居群间甚至居群内的核型存在巨大的变异,包括:(1)染色体数目变异很大,由16、17、18、19、21到24,并首次报道了三倍体居群(皇莆山细胞型Ⅰ);(2)T(端部着丝点染色体)存在一些异常类型,包括m’(小m型染色体)、sm(近中部着丝点区染色体)、st(近端部着丝点区染色体)、t(端部着丝点区染色体)、SAT(随体染色体)等类型;(3)核型类型大多为“3B”型,少数为“2B”、“2C”或“3C”型核型。对安徽省石蒜属两新记录种稻草石蒜和江苏石蒜的核型分析认为,两种均为杂交起源,稻草石蒜可能起源于中国石蒜和二倍体石蒜的杂交;江苏石蒜可能起源于二倍体石蒜和中国石蒜或长筒石蒜的杂交。4.研究了超声提取中国石蒜总生物碱的最佳工艺条件和不同时期总生物碱含量的动态变化,以及中国石蒜和石蒜不同产地、不同部位总生物碱含量。正交试验显示,在超声工作频率为40 KHz时,中国石蒜总生物碱最佳超声提取工艺为:提取温度60°C,乙醇浓度65 %,料液比为1 : 20,提取时间为2 h。中国石蒜和石蒜不同产地总生物碱含量存在较大差异;不同部位总碱含量均是果实最高,其次是花序,叶片中含量最低;其中,鳞茎中含量低于生殖器官。

【Abstract】 The species in Lycoris Herb. of Amaryllidaceae which are all typical bulbous geophytes with leaf appearing in autumn or spring, are endemic to East Asia and principally native to China, Japan and Korea. To date, studies of the genus Lycoris have focused mainly on morphology and anatomy, karyotypes, tissue culture and propagation, medicine and molecular aspects. Little is known about the leaf photosynthetic characteristics of Lycoris species. All species in Lycoris are traditional medicine, while the medicine elements of only a few species have been separated and identified. There are many prodigious disputes remained on the systematic and karyotypic studies of the genus Lycoris.This paper made detailed studies through measuring photosynthetic characteristics, analyzing karyotype and determining the content of total alkaloids on the materials including seven species of the genus Lycoris and different populations of L. radiata and L. chinensis from Anhui province. It could afford some basic scientific data for the discussion on the systematic ralationships and karyotype evolution of the genus Lycoris, and the further exploitation and comprehensive utilization of the Lycoris resources. Moreover, much difference was found not only in different Lycoris species, but also in different inter-populations and intra-populations of L. radiata and L. chinensis. The results were as follows:1. The diurnal changes of gas exchange and chlorophyll fluorescence parameters of four species with leaf appearing in autumn and three species with leaf appearing in spring in the genus Lycoris from Anhui province were determined and compared by a portable photosynthesis analysis system (CI-340). The results showed that marked difference existed in photosynthetic characteristics among different species; the other six species all exhibited a typical decline which was not observed in L. aurea in net photosynthesis rate (PN) during midday; all species had relatively low light saturation point (LSP), low light compensation point (LCP) and high apparent quantum yield (AQY), with the ranges being 322~1189μmol m-2 s-1, 8.11~15.36μmol m-2 s-1 and 0.0457~0.0798, respectively. It was supposed that the seven species were all typical shade plants.2. The effects of shading on the growth, leaf photosynthetic characteristics and chlorophyll fluorescence parameters of Lycoris radiata var. radiata (3n=33) were determined under differing irradiances. The plants experienced a pronounced depression in PN under full light at midday unlike those under shading condition. It was indicated that 15 % of ambient irradiance was too low and the open sky irradiance was too high for L. radiata var. radiata growth. When the integrated daytime carbon gain, leaf area and total leaf area per plant which are the main factors determining the productivity of L. radiata var. radiata plant were taken into account together, this species could be grown in moderate-shade environment with about 60~70 % of ambient irradiance to promote its growth.3. Great variations existed in the karyotype of the genus Lycoris, and a lot of new karyotypes and chromosome numbers were found by detailed cytological analysis. Tremendous inter-populational and intra-populational variations were also found in the karyotype of L. chinensis: (1) the variation of the chromosome numbers was very great, from 16, 17, 18, 19, 21 to 24; and one population of triploid (3n=24) cytotype (Huangpushan Population CytotypeⅠ) was first discovered here; (2) the terminal point chromosome (T) had diverse variations and some rearranged chromosomes which were aberrant had been found; the aberrants included small median centromeric chromosome (m’), submedian centromeric chromosome (sm), subterminal centromeric chromosome (st), terminal centromeric chromosome (t) and satellite chromosome (SAT); (3) Most of the karyotypes of L. chinensis were“3B”type, but there were also some“2B”,“2C”or“3C”types. The results of studying on the karyotypes of L. straminea and L. houdyshelii being new records in Anhui province showed that L. straminea probably derived from hybridization between L. chinensis and L. radiata var. pumila, and L. houdyshelii was possibly from hybridization between L. radiata var. pumila and L. chinensis or L. longituba.4. The best technological conditions for ultrasonic extraction of total alkaloids in L. chinensis were explored by one single factor and orthogonal experiments and the contents of that in different organs and in bulbs during different growth stages and from different places were determined. It was indicated that with the ultrasonic frequency being 40 KHz, the optimum technological conditions were: temperature at 60°C, the extraction reagent of 65% ethanol, material to extraction reagent 1 : 20, and the extraction time for 2 hours. There were significant differences in the content of total alkaloids among different places. The content of total alkaloids in fruit was the highest, followed by the inflorescence, with the lowest in leaf. The content of total alkaloids in bulb was lower than that in reproductive organs.

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