摘要
差不嘎蒿(A.halodendron Turcz.ex Bess)、褐沙蒿(A.intramongolica H.C.Fu)和黑沙蒿(A.ordosica Krasch.)均是菊科蒿属的半灌木,是干旱区重要的固沙植物,也是荒漠植被中的主要建群种和优势种,在荒漠区的生态环境保护中发挥着极其重要的作用。三种沙蒿亲缘相近,形态特征十分相似,对三种沙蒿间亲缘关系的研究,可更加深入地了解龙蒿组种间的亲缘演化关系,为其种间的分类提供更多分子水平上的依据。本文利用ISSR技术结合解剖特征及与环境因子的关系,对三种沙蒿进行研究,结果如下:
1.三种沙蒿均具有高的遗传多样性,不仅表现在物种水平而且表现在种群水平上。种群水平,多态位点百分率为褐沙蒿>黑沙蒿>差不嘎蒿,物种水平的顺序与种群水平一致,但三种沙蒿的值非常接近,褐沙蒿略高。从Nei基因多样度与香农多样性指数看,物种水平与种群水平均为差不嘎蒿>黑沙蒿>褐沙蒿,与多态位点百分率的顺序相反,说明差不嘎蒿的多态性低但均匀性高,褐沙蒿则相反。三种沙蒿的基因分化系数为差不嘎蒿Gst=0.1041、褐沙蒿Gst=0.0757、黑沙蒿Gst=0.0768,表明三个种均是绝大部分的遗传变异存在于种群内。
2.三种沙蒿17个地理种群的聚类分析显示,以种为单位聚为三大支,同一物种的各种群均能聚到一起,反映出三个种具有一定的分化。从种间关系来看,黑沙蒿5个种群构成的一大支首先与褐沙蒿的7个种群构成的大支相聚,然后再与差不嘎蒿5个种群的大支聚到一起,表现出黑沙蒿与褐沙蒿关系更为密切,物种水平上的聚类显示了同样的结果。
3.由解剖特征可知,三种沙蒿的叶及营养枝的解剖结构特征一致,反映了它们之间较近的亲缘关系。上下表皮的角质层均较厚,栅栏组织与海绵组织的比值较大,说明了三种沙蒿较强的抗旱能力。
4.差不嘎蒿、黑沙蒿的营养枝及叶解剖结构与环境因子的相关性分析显示,随着年降水量的降低,差不嘎蒿的叶上表皮角质层呈变厚的趋势,即叶上表皮角质层随干旱而变厚,以减少水分的散失。从叶下表皮角质层来看,差不嘎蒿随寒冷指数降低而变厚,黑沙蒿则随极低温的下降而增厚,表现出两种植物的叶下表皮角质层均随低温而增厚,叶厚也有同样的趋势,黑沙蒿叶厚随年均温的降低而增厚。差不嘎蒿栅栏组织厚度随着生长季降水量的减小而增大,髓茎比随着湿润指数的增大而增大,均体现出对水份的适应性。黑沙蒿营养枝基部直径则与最暖月温度有关,呈现正相关关系。再有,对于两种植物均表现出随温和度减小,营养枝皮层厚度增大,即表现出寒冷使皮层增厚。从决定系数(R2)来看,只有差不嘎蒿叶上表皮角质层与年降水、黑沙蒿营养枝基部直径与最暖月平均温度的R2值高,表明在这两个方面反映出的规律性强。
A. halodendron Turcz.ex Bess、A. intramongolica H.C.Fu and A. ordosica Krasch. are subshrub belonging to genus Artemisia of Compositae. They are edificators or dominant species of sand vegetation, important fixation plants and have played an important role in desert eco-environment protection. The three species have close kinships and quite similar morphological characteristics. The research of their kinships redounds to probe into the phylogeny of section dracunculus of genus Artemisia and provide more molecule-level proofs for the classification among the species. In this paper, the tree species were studied by ISSR makers being combined with anatomic characteristics and environmental factors, the results were as follows:
1. The three species had high genetic polymorphism in both species level or in population level. In population level, the sequence of polymoiphic loci percentage was: A. intramongolica > A. ordosica > A. halodendron. The sequence of species level was the same, but the values were very close to, only A. intramongolica slightly higher. Nei’s gene diversity index (H) and Shannon index(I) in both species level or in population level, the sequence all were: A. halodendron > A. ordosica > A. intramongolica, opposite to polymoiphic loci percentage. It indicated that A. halodendron had lower polymorphism and higher evenness, A. intramongolica was contrary. The genetic differentiation among populations (Gst) of the tree species were: A. halodendron Gst=0.1041, A. intramongolica Gst=0.0757, A. ordosica Gst=0.0768, it indicated more variations existed within the populations.
2. The cluster analysis on 17 geographical populations of the three species demonstrated that the populations of every species were clustered together respectively to three branches, it reflected the definite genetic differentiation among the species. To the relations of the species, the branch of five populations of A. ordosica were clustered together with the branch of seven populations of A. intramongolica firstly, and then with the populations of A. halodendron. It indicated that kinships were nearer between A. ordosica and A. intramongolica. The cluster analysis on the species level demonstrated the same results.
3. According to the anatomic characteristics,the leaves and foliage branch of the three species were coincident, reflected that they had quite close kinships. The thick upper and lower cuticle and the big ratio of the palisade mesophyll to spongy mesophyll indicated that the three species had strong drought-resistance ability.
4. Analyzing the correlation between the anatomic characteristics and environmental factors to the A. halodendron and A. ordosica , the result showed that: 1) With an decrease of annual precipitation, upper cuticle thickness of the A. halodendron increased, indicated the upper cuticle thicken with the drought to decrease the dissipation for water. 2) The lower cuticle thickness, both of the A. halodendron and A. ordosica tended to thicken with the temperature decrease. The leaf thickness of A. ordosica also had the same tendency. 3) The palisade mesophyll thickness of the A. halodendron tended to thicken with the growth season precipitation decrease and the ratio of medulla to stem increase with the wetness index increase, these reflected the adaptability to water. 4) It was the positive correlation between diameter of basal foliage branch of the A. ordosica and the mean temperature of the warmest month. 5) With the decrease of the temperateness, the cortex thickness of foliage branch of the two species thicken, indicated the coldness made cortex thicken. 6) According to the coefficients of determinant (R2), the value was high only in the upper cuticle thickness and the annual precipitation for the A. halodendron, the diameter of basal foliage branch and the mean temperature of the warmest month for the A. ordosica, it indicated the rule was obvious in the two aspect.
引文
[1]林有润.论世界蒿属植物区系[J].植物研究,1995,15(1):1-37.
[2]林有润.中国植物志[M].北京:科学出版社,1991:191-195.
[3]张德魁.两种蒿属植物的生理生态学特征及其防风固沙效应研究[D].甘肃农业大学,2006:
[4]林有润.中国蒿属志一中国蒿属植物的系统分类、分布和主要经济用途[M]. BULLETIN OF BOIANICAL RESEARCH. 1988,8(4):1-61.
[5]张燕.新疆五种蒿属植物的应用基础研究[D].西北师范大学,2004:
[6]富象乾.内蒙古植物志[M].第二版.呼和浩特:内蒙古人民出版社,1993: 641-643.
[7]王铁娟.中国北方六种沙蒿亲缘关系、替代分布规律与成因的研究[D].内蒙古大学,2004:
[8]林有润.蒿属的演化系统兼论蒿属与邻近属的亲缘关系[J].植物研究,1982,2(2):1-60.
[9]王铁娟,杨持,吕桂芬,等.中国北部六种沙蒿的地理替代规律及其主导生态因子[J].生态学报,2005,25(5):1012-1018.
[10]黄兆华,刘媖心.我国沙区重要蒿属植物的特性及应用[J].干旱区资源与环境,1991,5 (1):12-21.
[11]李进,张秀伏.差不嘎蒿特性的初步研究[J].中国沙漠,1991,11(1):50-58.
[12]张继,马君义,姚健,等.黑沙蒿资源的综合开发利用研究[J].中国野生植物资源, 2003,22(1):81-92.
[13]王立群,陈世璜,郝利忠.黑沙蒿生态生物学特性及群落地理分布规律相关性研究[J].干旱区资源与环境,2002,l6(4):95-98.
[14]杨洪晓,张金屯,吴波,等.油蒿对半干旱区沙地生境的适应及其生态作用[J].北京师范大学学报,2004,40(5):684-690.
[15]张德魁,王继和,马全林,等.油蒿研究综述[J].草业科学,2007,24(8):30-35.
[16]刘华民,朴顺姬,王立新,等.不同演替阶段褐沙蒿种群特征研究[J].生态学杂志, 2005,24(5):497-502.
[17]鲁作民.不同年龄油蒿抗旱性的研究[J].植物学通报,1993,10(1):58-59.
[18]龚吉蕊,张立新,赵爱芬.油蒿(Artemisia ordosica)抗旱生理生化特性研究初报[J].中国沙漠,2002,22(4):387-392.
[19]周海燕.水分胁迫对冷蒿和差不嘎蒿体内水分状况的影响[J].中国沙漠,1999,19(1):72-92.
[20]唐海萍,史培军.沙坡头地区不同配置格局油蒿和柠条水分生态位适宜度研究[J].植物生态学报,2001,25(1):6-10.
[21]李晓兵,杨劼,田青松.油蒿光合产物分配动态[J].中国草地,1996,2: 47-51.
[22]杨劼,高清竹,乌力吉,等.库布齐沙地油蒿(Artemisia ordosica )蒸腾作用特征及其与环境因子的关系[J].内蒙古大学学报(自然科学版),1999,30(3):372-376.
[23]朴顺姬,潮洛蒙,刘新文,等.不同类型沙漠化土地差不嘎蒿种群热值进行研究[J].中国沙漠,1999,19(1):82-87.
[24]黄振英,Yitzchak GUTTERMAN.油蒿与中国和以色列沙漠中的两种蒿属植物种子萌发策略的比较[J].植物学报,2000,42(11):71-80.
[25]刘芳,朴顺姬,刘华民.褐沙蒿种子对干热生境的适应性研究[J].水土保持通报,2008,28(2):19-29.
[26]李进.差不嘎蒿的分布及其在天然植被演替中的地位[J].中国沙漠,1991,11(2):55-60.
[27]常学礼,赵学勇,李胜功,等.差不嘎蒿在科尔沁沙地草场植被中的作用[J].中国草地,1996, 16(1):27-31.
[28]崔望诚.半荒漠地带的油蒿群落[J].中国草地,1991,2:50-52.
[29]王庆锁,李博.鄂尔多斯沙地油蒿群落主物量初步研究[J].植物生态学报,1994,18 (4):347-353.
[30]王庆锁.油蒿、中间锦鸡儿生物量估测模式[J].中国草地,1994,1:49-51.
[31]王庆锁,粱艳英.油蒿群落植物多样性动态[J].中国沙漠,1997,17(2):159-163.
[32]王庆锁,董学军,陈旭东.油蒿群落不同演替阶段某些群落特征的研究[J].植物生态学报,1997.21(6):531-538.
[33]潮洛蒙,朴顺姬,智瑞年,等.不同类型沙地中差不嘎蒿种群分布格局进行研究[J].中国沙漠,1999,19(1):54-84.
[34]郭柯.毛乌素沙地油蒿群落的循环演替[J].植物生态学报,2000,24(2):243-247.
[35]郭柯,董学军,刘志茂.毛乌素沙地沙丘土壤含水量特点——兼论老固定沙地上油蒿衰退原因[J].植物生态学报,2000,24(3):275-279.
[36]张德魁,王继和,马全林,等.腾格里沙漠南缘油蒿与沙蒿种群分布格局[J].甘肃科技,2008,24(3):721-921.
[37]王凌诗,邢淑清,张贵一.中国东北蒿属叶表皮特征研究I—龙蒿组的研究[J].哈尔滨师范大学自然科学学报,1994,10(2):90-97.
[38]邢怡,张大维,宫晓波,等.东北地区蒿属植物比较形态学研究Ⅱ.叶表皮结构[J].植物研究,2000,20(1):48-57.
[39]蒋林,林有润.中国蒿属比较形态和解剖学研究Ⅲ茎、叶解剖结构[J].仲恺农业技术学院学报,1997,10(1):12-23.
[40]山宝琴,贺学礼.毛乌素沙地12种蒿属植物叶的解剖特征[J].西北农林科技大学学报(自然科学版),2007,35(6):211-217.
[41]张金谈,王萍莉,郝海平,等.现代花粉形态研究[M].北京:科学出版社,1990,26-51.
[42]蒋林,林有润.中国蒿属比较形态和解剖学研究Ⅱ[J].花粉形态,1996,4(3):1-14.
[43]王铁娟,张颖娟.六种沙蒿花粉形态的比较研究[J].内蒙古师范大学学报(自然科学汉文版),2006,35(1):83-88.
[44]马骥,李俊祯,孔红.我国沙区6种蒿属植物的种子微形态特征[J].中国沙漠,2002, 22 (6):586-590.
[45]鲁作民.油蒿和籽蒿种子化学组成的研究[J].植物学通报,1995,12(1):54-55.
[46]张继,马君义,姚健,等.黑沙蒿资源的综合开发利用研究[J].中国野生植物资源, 2003,22(1):27-29.
[47]颜世芬,梁冰,李菊白,等.黑沙蒿微量元素及精油的化学成分研究[J].分析实验室,1994,13(3):82-84.
[48]于凤兰,马茂华,孔令韶.油蒿挥发油的化感作用研究[J].植物生态学,1999,23(4):345-350.
[49]赵东保,杨玉霞,张卫,等.黑沙蒿黄酮类化学成分研究[J].中国中药杂志,2005, 30(18):1830-1834.
[50]李明静,张卫,赵东保,等.三波长-分光光度法测定黑沙蒿中的总黄酮[J].分析实验室,2007,26(3):99-102.
[51]乔有明,富象乾,阎贵兴,等.20种蒿属植物的染色体核型研究[J].中国草地,1990, 6: 24-31.
[52]曹瑞,张寿洲.两种蒿属植物的染色体数目和核型研究[J].武汉植物学研,1990, 8(1):9-12.
[53]扈廷茂,曹瑞,王汉屏,等.内蒙古五种蒿属植物的染色体研究[J].内蒙古大学学报(自然科学版),1991,22(3):422-427.
[54]王凌诗.中国东北蒿属龙蒿组植物核型研究[J].植物研究,2000,20(4):403-410.
[55]王铁娟,杨持,尹俊.内蒙古蒿属沙生地理替代种的ITS序列的比较分析[J].内蒙古大学学报(自然科学版),2004,35(5):550-556.
[56]王铁娟,王静,吴恩岐.褐沙蒿遗传分化的研究[J].内蒙古师范大学学报(自然科学汉文版),2004,33(4):428-431.
[57]王铁娟,杨持,马静,等.油蒿种群遗传分化的RAPD分析[J].内蒙古大学学报(自然科学版), 2004,35(4):399-403.
[58]王铁娟,韩国栋.中国蒿属沙生地理替代种亲缘关系的RAPD分析[J].植物研究,2007, 27(4):449-454.
[59] Bostein D R, White R L, Skolnick M et al. Am J Hum Genet, 1980, 32: 314-331.
[60] Shoemaker R C, Specht J E. Integration of the soybean molecular and classical genetic-linkage groups[J]. CropScience, 1995, 35: 436-446.
[61] Szmidz, A. E. and Wang X. R.. DNA markers in forest genetics. In Mu1lerStarch G. and M.Ziehe (eds.). Genetic Variation in European Populations of Forest Tree [M]. Frankfurt:Sauerlander's Verlag, 1991,79-91.
[62]郭大龙,罗正荣.一种利用ISSR开发SSR引物的方法[J].植物生理学通讯,2006,42(2):268-270.
[63] Queller D. C, Strassmann J. E. and Hughes C. E. Microsatellites and kinship [J]. Thends Ecol.Evo1, 1993, 8: 285-288.
[64]张军丽,土峥峰,李鸣光,等.植物种群研究中的分子标记及其应用[[J].应用生态学报, 2000, 11(4): 631-636.
[65] Williams JG, Kubelik AR, Livak KJ, et al. DNA polymorphisms amplified by arbitrary primers are useful genetics markers[M]. Nucleic Acids Research, 1990, 18: 6531-6535.
[66] Welsh J and McClelland M. Fingerprinting genomes using PCR with arbitrary primers[M]. Nucleic Acids Research, 1990, 18: 7213-7219.
[67]邹喻萍,葛颂,王晓东.系统进化植物学中的分子标记[M].科学出版社,2001, 1: 30-51.
[68] Zietkiewicz E, Rafalski A. and Labuda D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification[J]. Genomics, 1994, 20: 176-183.
[69]聂书萍,代林远.分子标记的发展及应用[J].河南职工医学院学报,2006,18(6):526-528.
[70] Esselman E. J., Li J. Q., Crawford D. J., Windus J. L., and Wolfe A. D. Clonal diversity in the rare Calamagrostis porteri ssp. Insperata (Poaceae): comparative results for allozymes and random amplified polymorphic DNA(RAPD) and intersimple sequence repeat(ISSR)markers[J]. Molecular Ecology, 1999, 8: 443-443.
[71]袁昭岚,沈颂东,黄鹤忠,等.SSR和ISSR分子标记技术及其在遗传多态性方面的应用[J].水产养殖,2005,26(2):10-13.
[72]刘木根,赵寿元.单核苷酸多态性概述[J].生命科学,2000,12 (6):277-281.
[73]朱颜,周国利,吴玉厚,等.遗传标记的研究进展和应用[J].延边大学农学学报,2004,26(1):64-69
[74] Zabeau M, Vos P, Kuiper M et al. Combined mapping of AFLP and RFLP markers in barley [J]. Mol. Gen. Genet, 1995, 249: 65-73.
[75]陈婴芳,徐宏发.DNA分子标记及其在保护生物学中的应用[J].浙江师范大学学报(自然科学版),2003,26(1):60-64.
[76]张传军,刘亦肖,肖娅萍.遗传多样性与植物的遗传标记[J].陕西师范大学学报(自然科学版),2006,34:275-278.
[77] Weising K, Nybom H, Wolff K, Meyer W. DNA fingerprinting in plants and fungi[M].CRC Press, Inc., Bota Raton, 1995.
[78]王中仁.植物遗传多样性和系统学研究中的等位酶分析[J].生物多样性,1994,2(2):91-95.
[79] Schaal B. A., Hayworth D. A.,Olsen K.M., Rauscher J. T.,Smith W. A.. Phylogeographic studies in plants: problems and prospects [J]. Molecular Ecology, 1998,7(4):456-474.
[80] Anamthawat KJ.,Bragason BTH, Bodvarsdottir SK, Koebner RMD. Molecular variation in Leymus species and populations [J]. Molecular Ecology,1999,8(2)::309-315.
[81] Pelah D, Wang WX, Altman A. Differential accumulation of water stress-related proteins,sucrose synthase and soluble sugars in Populus species that differ in their water stess response[J]. Physilogia Plantarum,1997,99:153-159.
[82] Ryan KO,Baum DA. Phylogenetic relationships of North American Antirrhinum (Veronicaceae) [J]. American Journal of Botany, 2004, 91:918-925.
[83] Nei M. Molecular population genetics and evolution [M].Amsterdam: North Holland Co, 1975.
[84] King, L. M., B. A. Schaal.. Ribosomal DNA variation and distribution in Rudbeckia missouriersis [J]. Evolution,1989, 43:1117-1119.
[85] Nei M. Analysis of gene diversity in subdivided population [J]. Proc Nat Acad Sci USA,1973, 70:3321-3323.
[86] Nei M. Genetic distance between populations [J]. American Naturalist, 1972, 106: 283-292.
[87]何凤仙.植物学实验[M].北京:高等教育出版社,2000:18-23.
[88]贺学礼.植物学实验实习指导[M].北京:高等教育出版社,2004:26-34.
[89] Kira T. A new classification of climate in eastern Asia as the basis for agricultural geography[M]. Kyoto: Kyoto University (Horticultural Institute) Press, 1945.
[90] Kira T. On the altitude arrangement of climatic zone in Japan [J]. Kanchi Nougaku, 1948.2:143- 173.
[91] Holdridge L R. Life Zone Ecology [M]. Cost Rica: San Jose Press, 1967.
[92] Bailey H P. The mean anna range and standard deviation as measures of dispersion of temprature around the annual mean [J]. Geogrofiska Annaler, 1966, 48: 183-194.
[93]徐文铎,东北地带性植被建群种及常见种的分布与水热关系的研究[J].植物学报,1983,25 (3):264-274.
[94] Bailey H.P. Semi-arid climates: their definition and distribution[J]. Ecol Study, 1973, 34: 73-97.
[95]钱韦,葛颂.局群遗传结构研究中显性标记数据分析方法初探[J].遗传学报,2001,28 (3):244-255.
[96]费松林,方精云,樊拥军等.贵州梵净山亮叶水青冈叶片和木材的解剖学特征及其与生态因子的关系[J].植物学报,1999,41(9):1002-1009.
[97]张新英,曹宛红.生长在不同生境下的沙棘次生木质部解剖学的研究[J].植物学报(英文版),1990,32:909-915.
[98] Yuan Y. M (袁永明),Peng Z. X(彭泽祥). The systematic and ecological significance of anatomical characters of leaves in the tribe Thermopsideae (Fabaceae)[J]. Acta Bot Sin(植物学报),1991,33:840 847 (in Chinese).
[99] Donselman HM , Flint HL. Genecology of eastern redbud [J]. Ecology , 1982,63 (4) :962–971.
[100]王勋陵,王静.植物的形态结构与环境[M].兰州:兰州人学出版社,1989: 105-138.
[101]朱栗琼,李吉跃,招礼军.六种阔叶树叶片解剖结构特征及其耐旱性比较[J].广西植物,2007, 27(3):431-434.
[102] Levittt J . Response of plants to environmental stress [M]. New York :Academic press,1972.
[103] JACKSON L W R. Effect of shade on leaf structure of deciduous tree species [J]. Ecology. 1967, 48:498-499.
[104] HUNER N P A,PALTA J P,LI P H,et al. Anatomical changes in leaves of Pumarye in response to growth at cold- hardening temperature[J]. Bot. Gaz.,1981,142:55-62.
[105]李正理,李荣敖.1981.我国甘肃九种旱生植物同化枝的解剖观察[J].植物学报,23(3):181-185.