用户名: 密码: 验证码:
盾叶薯蓣遗传多样性及多倍体新种质培育的研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
我国特有种盾叶薯蓣(Dioscorea zingiberensis C.H.Wright)根茎中薯蓣皂苷元含量高,是提取甾体激素类药物的重要药源植物。目前,栽培种源混乱,种质退化,低含、低产,遗传背景不清,缺乏优良品种。为此,本研究从其资源调查入手,利用RAPD技术对其遗传多样性及系统分类学进行了研究。并通过倍性诱变技术,培育出高含、高产、遗传性状稳定的同源四倍体新种质XHJ—401,建立了其离体快繁技术体系,对其生物学与农艺学特性及生理生化特性进行了研究,并与AK杂交获得了杂交种子。主要研究结果叙述如下:
     (1)以TY雄株为起始材料,通过离体化学诱变获得了同源四倍体染色体为2n=2x=40。其一、二年生试管苗根茎平均产量分别为7.87725×10~3kg·hm~(-2)和8.70558×10~4kg·hm~(-2),较其2n试管苗(2N)分别高141%和373.11%;一、二年生根茎薯蓣皂苷元平均含量分别为1.601%和2.966%,后者较2N和TY分别高255.21%和81.30%;其光合作用、呼吸作用、叶片叶绿素含量、气孔器及保卫细胞大小、保卫细胞叶绿体数量及大小、POD及CAT活性均高于2n亲本,田间的适应性及营养生长也强于2n,生育期较之长37~40d,达306d~310d;性状稳定;具有高产及高抗的形态结构与生理生化基础。与AK杂交,获得杂交种子的染色体为2n=3x=30;花药培养所获愈伤的染色体为n=1x=10。
     (2)对其形态学多样性进行研究。发现了自然六倍体(2n=6x=60)和四倍体(2n=4x=40);首次发现盾叶薯蓣有多胚现象,可能具有无融合生殖;发现有雌雄异株、雌雄同株或同序、杂性同株或异株及具不育雄蕊的两性花;自然性比:雄株:雌株:雌雄同株及同序=54:44:2;发现了块茎状根状茎、长棍状根状茎,异形叶性,叶脉凹陷型叶片;观察到其花序是莱荑花序而不是穗状花序;各种群器官形态变化因种群、个体、所处部位及立地环境不同而存在明显差异。
     (3)对其生物学和农艺学特性进行研究时发现,其雄株、雌株、雌雄同株三者可以自然相互转换,变性率:XHJ-401(12.69%)>2N(10.83%)>TY(10%);具有单性结实现象,结实率:XHJ-401(36.4%)>AK(8.2%)。具耐荫植物特点。薯蓣皂苷元含量与多个形态指标具有相关;根状茎断面颜色不能作为含量的形态标记,但在同一种质之内比较可行;根据保卫细胞宽度能快速准确地进行倍性形态学鉴定。
     (4)对盾叶薯蓣种群进行了RAPD遗传多样性、遗传结构的比较研究,提出盾叶薯蓣为遗传多样性较高、Gst相对偏小而Nm相对较大的亚狭域种。主要结果如下:①从60条随机引物中筛选出条带清晰可辨、多态性表现较为丰富的20个引物,对52份供试材料进行RAPD PCR扩增,共扩增出385条DNA带,其中220条为多态带,PPB=57.14%。②利用POPGENE 1.32软件进行遗传多样性分析表明,盾叶薯蓣的Nei’s h=0.4737,Shannon I=0.6666,具较大的遗传多样性;在薯蓣属6个种中的遗传多样性最高。其遗传多样性大小及其顺序为:种间(h=0.4767,I=0.6697)≥种内≥种群间(0.4737,0.6666)≥种群内(0.4328~0.4829,0.6310~0.6758)≥世代间的(0.4500,0.6414)≥亲本世代(AK:0.4496,0.6410)≥F1代的(AK F1:0.4409,0.6308)。种群内遗传多样性相对丰富,但各种群的遗传多样性存在明显差异,江南种群>江北种群。③种间Gst=0.0709>种群间Gst(0.0390)>世代间Gst(0.0075)。薯蓣属种内分子变异大种间分子变异小;盾叶薯蓣种群内、世代内的分化大,种群之间、世代之间的分化小。④盾叶薯蓣世代间Nm(66.5163)>种群间Nm(12.3071>种间Nm(6.5516);其种群间Nm大,但不同种群Nm有所不同,差别较大。⑤Nei's遗传距离D值大小及顺序为:纤细薯蓣(0.0289)<叉蕊薯蓣(0.0612)<柴黄姜(0.0651)<薯蓣(0.0733)<黄独(0.0782);盾叶薯蓣各种群间D=0.0067~0.1575。薯蓣属3组6种之间:D=0.0289~0.1105,遗传一致度I=0.9248~0.9715;根状茎组3种之间的D值最小,与基生翅组黄独间的D值最大。XHJ-401与2N及TY亲本之间的D值分别为0.0350和0.0333,发生了较大的遗传分异。
     (5)盾叶薯蓣种群UPGMA聚类分析结果显示,染色体倍性相同,则遗传距离相近,因而来源不同但倍性相同的类群能较早聚为一类,构成种群复合体(新提概念)。倍性变化产生的遗传分异大于地理隔离和种群扩散距离,高山的遗传隔离作用较江河的大。染色体加倍,造成了形态学表型及NDA分子的较大差异,因而提出其多倍体生物学效应并非来源于简单的DNA量的累加效应的观点。
     (6)建立了盾叶薯蓣种内分子分类系统,提出了根状茎组→周生翅组→基生翅组薯蓣属组间进化新观点;首次以分子证据证明了根状茎组为一个自然类群。
     (7)绘制了盾叶薯蓣资源水平分布图,发现其两个主要分布区并不明显连续;认为其种群扩散路线是沿长江及其支流由西向东、并在长江中游再分支向南、向北迁移,形成以湘西和鄂西北为中心的两个新的分布亚中心。提出其种群扩散的主要阻遏因素不是大江而是高山的新观点。本种分布于多种乔木林、灌丛及草丛群落中;自然分布率:灌丛(50.2%)>乔木林(34.6%)>草丛(15.2%);郁闭度3~5时的分布率分别高达48.4%、55.6%、45.2%。
Dioscorea zingiberensis C. H. Wright is an endemic plant of China, and is alsoan important medicine material plant for steroid extracting for its high contents ofdiosgenin in its rhizome. But the germplasms have degenerated, lower content ofdiosgenin and lower production of rhizome by cultivation because of its confusionpropagation seedlings. We have no much knowledge about the genetic background.And there are any high quality breed at present. The resources investigation has beenmade firstly, then the Genetic diversity and Systematics by RAPD markers have beenstudied by author. A new germplasm "XHJ-401"(autotetraploid D. zingiberensis)with high content of diosgenin and high yield of rhizome has been created bychemical mutagen with itro culture; and the technique system of its mass clonalpropagation has also been established; its agronomic, biological and biochemicalcharacteristics have been studied; the triploid hybrid seeds from XHJ-401×AK havebeen obtained. The main results are as fellows:
     (1) An autotetraploid XHJ-401 (2n=4x=40) has been created through chemicalmutagen with itro culture from male of population TY. Its average output of rhizomeof the annual and biennial test-tube plantlets is at 7.87725×10~3 kg·hm~(-2) and8.70558×10~4 kg·hm~(-2) respectively, 141% and 373.11% higher than that of the biennialdiploid parents respectively; and its average diosgenin content is at 1.601% and 2.966% respectively; the latter is 255.21% and 81.30% higher than that of 2N and TYrespectively. It is higher or larger in photosynthesis rate, stomatic conductance,transpiration rate, content of chlorophyll a & b, Peroxidase activities, Catalaseactivities, size of stoma & guard cell, number & size of chloroplast in guard cell ofleaf than that in its 2n parents; both the adaptability in field and the growth conditionare better than that of the diploid. It lasts 306d~310d in period of growth anddevelopment, 37d~40d longer than that of the latter. Its inherited properties are stabile;and its high yield and stress resistance come from its special morphological andstructural organs, biological & biochemical characteristics. The chromosome numberof the hybrid from XHJ-401×AK is 2n=3x=30; and the pollen callus is n=1x=10.
     (2) The morphological diversity of D. zingiberensis has been studied. The naturalhexaploid and two tetraploids was found. The apomixes is possible in D. zingiberensis because the polyembryony has been found firstly. Beside the dioecious andmonoecious, the polygamomonoecious, polygamo-dioecious, hermaphrodite flowerwith sterility stamen, staminate flower and pistillate flower in the same inflorescencehad also been found. The sexual rate in nature is males: females: monoecious=55: 44:2. Tuberous rhizome, long clavate rhizome, heterophylly, vein depressed leaf was alsofound, The inflorescence of this species is catkin but spike. The organ morphologicdifferences changes in different populations, bodies, positions in plant, and habitats.
     (3) The male, female and monoecious plant of this species can be denaturalizedand changed each other; The denaturalization rate of XHJ-401, 2N & TY is 12.69%,10.83% & 10% respectively. The parthen carpy be approved, and the fruiting rate ofXHJ-401 (36.4%) is much more than that of AK (8.2%). It is a shade-enduring plant.The contents of diosgenln in rhizome is correlated with a few morphologic characters.We can use the color to estimate the diosgenin content only in the same germplasm.The width of guard cell can be used to judge the ploidy more credibly.
     (4) Comparative studies on the genetic diversity, population genetic structure ofD. zingiberensis have been made. It has been pointed that this species is a narrowspecies with higher genetic diversity, lower coefficient of gene differentiation (Gst)among populations, and higher estimate of gene flow (Nm). The main results are asfellows:①20 RAPD primers from 60 10-base arbitrary primers have been chosenfor their strong, reproducible amplification and distinct polymorphisms. 385 loci wereidentified with this oligonucleotide primers, out of which 220 loci were polymorphicand account for 57.14% of total genetic diversity at species level. The result showsthat the materials tested have a high level genetic diversity.②Analysis of geneticdiversity with POPGENE shows the genetic diversity of D. zingiberensis is higherthan that of other 5 species in the same genus; The Shannon's indices of diversity (I)and Nei's gene diversity (h) is 0.4737 & 0.6666 respectively. The genetic diversityamong species (h=0.4767, I=0.6697)=that within species (0.4737, 0.6666)=thatamong populations (0.4737, 0.6666)=that within populations=that amonggenerations (0.4500, 0.6414)=that of parents (AK: 0.4496, 0.6410)=that of F_1(AKF_1: 0.4409, 0.6308). The genetic diversity within populations is abundant; but it isobviously different in different populations. The genetic diversity of populations inthe south side of the Yangtsc Rive is larger than that in the north side of the river.③The Gst among species is 0.0709, larger than that among populations (0.0390) andthat among generations (0.0075). This means molecular variations exist mostly withinspecies but beteen species as for dioscorea; and mostly within populations but beteen populations as for D. zingeberensis.④The Nm among generations is 66.5163,larger than that among populations (12.3071), and much larger than that amongspecies (6.5516). The Nm of populations is large, but is different from population topopulation.⑤The analyses of Nei's genetic distance (D) between every every twospecies of Discorea showed that the value of D is D. gracillima(0.0289)<D. Colletti(0.0612)<D. nippanica subsp, rosthornii(0.0651)<D. opposita (0.0733)<D. bulbifera (0.0782); The D value of every two populations of D. zingiberensis is0.0067(AK-SM)~0.1575(TJ-SZH); Among 6 species: D=0.0289~0.1105, I(genetic identity)=0.9248~0.9715; D value of 3 species of Sect. Stenophora is theclosest, and Sect. Opsophyton (D. bulbifera) is the farthest. The D value betweenXHJ-401 and its parents (2N & TY) is 0.0350 & 0.0333 respectively; This means thatbig genetic variation has occurred between tetraplois XHJ-401 and its diploid parents.
     (5) UPGMA Clustering analysis to populations of D. zingiberensis shows thatthe taxa with same ploid has similar genetic distance; So, they can be grouped into asingle group early, this is called "population complex" (A new conception). The effectof ploid on genetic variation is much more larger than that of geographical obstructand spread distance; mountain' much more than fiver'. The differences of phenotypeand DNA can be caused by ploid doubled. So the biological effect of polyploid is notonly caused by accumulation effect of DNA, but also by other factors.
     (6) Molecular systematics system of taxa under species of D. zingiberensis hasbeen established. It shows that Sect. Stenophora is a natural taxon; and their evolutionsequence is from Section Stenophora to Enantiophyllum then to Opsophyton.
     (7) The geographical distribution map of D. zingiberensis has been ploted. Itshows the two main distribution area are not connected distinctly. So, the populationdiffusion is possibly along the lines of the Yangtse Rive and its branches from west toeast, and then branching to north and south respectively in the middle drainage area ofthe rive, and forming two subcentre, that is west Hunan and northwest Hubei. Thehigh mountains is the main obstruct factor for its population spread. D. zingiberensiscan distribute in three kinds of communities including forests, shrubs and tussock; andits natural distribution rate is at 50.2%, 34.6% and 15.2% respectively; and 48.4%、55.6%、45.2% respectively when the Canopy density is at 3~5.
引文
1.中国植物志编委会.中国植物志(第十六卷第一分册).北京:科学出版社.1985:60~105.
    2.中国科学院植物研究所主编.中国高等植物图鉴(第5册).北京:科学出版社,1983,557~579.
    3.徐成基.中国薯蓣资源.成都:四川科学技术出版社,2000.
    4.丁志遵,唐世蓉,秦慧贞,等.甾体激素药源植物.北京:科学出版社,1983.
    5.崔天义,张丽红,糜留西,等.盾叶薯蓣灭钉螺活性成分的研究.武汉植物学研究,1998,16(1):65~68.
    6.江苏新医学院.中药大辞典(上册).北京:人民卫生出版社,1985:166~168.
    7.刘承来,陈延镛,唐易芳,等.盾叶薯蓣中甾体皂甙的分离和鉴定.植物学报,1984,26(3):283~289.
    8.李忌,陈俊杰,巨勇,等.天然甾体皂甙化合物的抗肿瘤活性.天然产物研究与开发,1999,11(1):14~17.
    9.刘乘来,陈延镛.小花盾叶薯蓣中甾体皂甙的分离和鉴定.植物学报,1985,27(6):635~639.
    10.刘乘来,陈延镛.薯蓣属植物的化学成分的研究Ⅱ叉蕊薯蓣中甾体皂甙的分离和鉴定.药学学报,1983,18:597~606.
    11.刘承来.从低熔点的薯蓣皂甙元中分离丝兰皂甙酮.医药工业,1983,(3):1~3.
    12.秦天才,张友德,张君芝.湖北黄姜资源的现状及开发利用.自然资源.1996,3:58~61.
    13.秦天才,张友德,张君芝.重要的甾体激素药源—黄姜.植物杂志.1997,1:8.
    14.国家医药管理局医药工业情报中心站.全国有机药物中间体索引,1989,497~559.
    15.L.特雷格著,邹继超译.类固醇激素.北京:科学出版社,1980,4~14.
    16. K. Takeda. The Steroidal Sapogenins of the Dioscoreaceae. Progress in Phytochemistry. 1972, 3: 288~289.
    17.唐世蓉,吴余芬,庞自洁.盾叶薯蓣甾体皂苷的分离与鉴定.植物学报.1983,25(6):556~561.
    18. Knuth, R. Dioscoreaceae. In Engl. Das Pflanzenreieh. 1924, 87(Ⅳ. 43): 1~387.
    19.中国科学院昆明植物研究所.云南植物志(第三卷)M.北京:科学出版社.1983:711~747.
    20.丁志遵.中国薯蓣科新分类群.植物分类学报,1982,20(2):205~209.
    21.万金荣.叉蕊薯蓣类群分类学的初步研究.南京中山植物园研究论文集.南京:江苏科学技术出版社,1990.41~44.
    22. Prain, D. & Burkill, I. H. Chromosome Numbers and Behavior in Some Species of Dioscorea, Cytologia, Tokyo. 1963, 28: 96~101.
    23. Burkill, I. H. The Organography and the Evolution of Dioseoreaceae, the family of the Yams, J. Linn. Soc. (Bot). 1960, 56: 319~416.
    24. Burkill, I. H. Ann. Royal Bot. Gdn., Calcutta, 14. Part. Ⅱ. 1938, Plates 86~146.
    25.秦慧贞,李碧嫒,吴竹君.盾叶薯蓣的胚胎发育及其在演化上的意义.中国植物学会五十五周年年会学术论文摘要汇编.中国植物学会.1988,307~308.
    26.江苏植物研究所薯蓣课题组.中国薯蓣属根茎组植物的分类和染色体数的研究.植物分类学报,1976,17(3):61~71.
    27.裴鉴,丁志遵,秦慧贞,等.中国薯蓣属根状茎组系统分类的初步研究.植物分类学报,1979,14 (1):65~-72.
    28.秦慧贞,张美珍,凌平平,等.中国薯蓣属细胞分类的研究—染色体数与该属起源和演化.植物分类学报,1985,23(1):11~18.
    29.魏宏图,丁志遵,徐克学.应用群分析研究薯蓣的分类.云南植物研究,1983,5(3):231~237.
    30.奚镜清,毛宗秀,杨建伟,等.浙江省薯蓣科植物的数量分类研究.植物分类学报,1990,28(6):442~451.
    31.凌萍萍.薯蓣属(Dioscorea)叶表皮气孔类型上的意义.南京中山植物研究论文集.南京:江苏科学技术出版社,1982.11~16.
    32.凌萍萍.薯蓣属(Dioscorea L.)植物的过氧化物同工酶及其在分类上的意义.南京中山植物园研究论文集.南京:江苏科学技术出版社1985,19~23.
    33.张美珍.薯蓣属(Dioscorea L.)茎的比较解剖及在分组上的意义.南京中山植物园研究论文集.南京:江苏科学技术出版社,1982.1~10.
    34. Ding Z. Z, Gilbert M G and Turland N. A new name in Chinese Dioscorea (Dioscoreaceae). Novon, 2000,10(1): 13.
    35. Martin F W, Delpin H. Techniques and problems in the propagation of sapogenin-bearing yams from cuttings. Agri Univ Puerto Rico, 1969, 53: 191~198.
    36.奚元龄,颜昌敬.植物细胞培养手册.北京:农业出版社,1992,408~415.
    37. D. Lauzaer, G. Laublin et al. In vitro propagation and cytology in wild yams, D.abyssinica Hoch. and D. mangenotuana Miege. Plant Cell, Tissue and Organ Culture. 1992, 28: 215~223.
    38.黄涛,张友德,张君,秦天才.黄姜种内染色体数目的变异.华中农业大学学报,2002,21(2):158~160.
    39.郑晓琴,梁国鲁,李晓林.盾叶薯蓣自然变异类型间的比较研究.亚热带植物学报,2003,43.
    40.李光明.盾叶薯蓣离体培养技术的优化及其同源四倍体的诱导.湖南农业大学硕士论文,2002.6.
    41.李运合.盾叶薯蓣四倍体诱导及快速繁殖技术的研究.华中农业大学硕士论文.2003.3.11(3):267~170.
    42. Jayanti Sengupta et al. Organogenesis and tuberization in cultures of D. floribunda. Plant Cell, Tissue. and Organ Culture. 1984, 3: 325~331.
    43. Martine Jean & Mario Cappadocia, In vitro tuberization in Dioscorea alata L. 'Brazo fuerte' and 'Florido' and D. abyssinica Hoch. Plant Cell, Tissue and Organ Culture. 1991, 26: 147~152.
    44. Bernard Malaurie et al. Effect of growth regulators concerntrations on morphological development of medstem tip in D. cayenensis, D .rotundata complex and D. praehensilis. Plant Cell, Tissue and Organ Culture. 1995, 41: 229~235.
    45. Shou Yong Choy NG. In vitro tubedzation in white yam (D. rotundata Poir). Plant Cell, Tissue and Organ Culture. 1988, 14: 121~128.
    46.四川省生物研究所一室体细胞组.盾叶薯蓣组织培养研究初报.植物学报,1978,20(3):279-280.
    47.江苏植物研究所.盾叶薯蓣试管植株的诱导.植物生理学通讯,1985,5:31.
    48.任建伟,白云.盾叶薯蓣愈伤组织诱导及培养.中国药学杂志,1993,28(9):532-534.
    49.任建伟,白云.盾叶薯蓣培养细胞的生长及薯蓣皂甙元产生的变化规律.中草药,1994,25(2):93~94.
    50.任建伟,白云.盾叶薯蓣悬浮培养细胞固定化的初步研究.中国中药杂志,1994,19(9):529~531.
    51.张宗勤,撒文清,刘建才.叉蕊薯蓣的微繁殖及微型薯蓣的离体诱导.生物技术.1998,8(1):18~20.
    52.江天生,李献军.盾叶薯蓣的组织培养及其废液利用初探.吉首大学学报,1999,20(1):85~87.
    53.谢碧霞,何业华.盾叶薯蓣愈伤组织培养及其高产系的筛选.中南林学院学报,1999,19(4):17~21.
    54.孟玲,朱宏涛.盾叶薯蓣的快速繁殖.天然产物研究与开发,2000,12(6):17~21.
    55.徐向丽.盾叶薯蓣的快速繁殖及化学诱变技术的研究.湖南农业大学硕士论文,1999.7.
    56.徐向丽,朱至清.盾叶薯蓣离体培养及微块茎的离体诱导.湖南农业大学学报,2000,26(4):282~285.
    57.徐向丽.薯蓣植物组织培养研究进展.湖南林业科技,2000,27(1):5~9.
    58.王志安,王生君.诱导盾叶薯蓣四倍体的研究初报.中国中药杂志.1995,20(6):337-339.
    59.吴玮,贾春,刘兴剑.盾叶薯蓣组织培养和植株再生.中国医学生物技术应用杂志.2004,4:41~43.
    60.易志军.盾叶薯蓣愈伤组织培养研究.经济林研究.2001,9(3):21~22.
    61.陈永勤,樊晋宇,易飞,等.盾叶薯蓣成熟叶片植株再生的研究.中国中药杂志.2004,29 (2):129~131.
    62.刘艳丽,姚家玲,张友德.盾叶薯蓣属伤组织诱导研究.华中农业大学学报,2004,23(4):389~392.
    63.莫兰,兰利琼,卿人韦,等.盾叶薯蓣种子萌发条件及诱导外植体愈伤的研究.四川大学学报,2004,41(4):837~841.
    64.李文谦,袁丽红,顾永明,等.盾叶薯蓣细胞生长特征的研究.南京工业大学学报,2004,26(5):12~16.
    65.彭菲,刘军,肖艳.不同外植体和激素组合对盾叶薯蓣组织培养的影响.湖南中医学院学报,2004,12(6):12-13,17.
    66.宋为民,陈为民.盾叶薯蓣试管植株的诱导.植物生理学通讯,1981,(5):31~37.
    67.晏婴才,林宏辉,代其林,等.盾叶薯蓣组织培养与快速繁殖研究.四川大学学报,2002,39(1):136~140.
    68.李涵,吴福川,郑思乡.盾叶薯蓣组织培养基四倍体诱导技术的初步研究.中国农学通报.2004,20(3):33~35.
    69.毕世荣,张忠福,苏成端.高含量薯蓣皂素植株的细胞克隆.天然产物研究与开发,1997,9 (4):1~6.
    70.李明芳.盾叶薯蓣愈伤组织与悬浮细胞的培养研究.湖南农业大学硕士论文,2000,7.
    71.李明芳,刘选明,刘斌,等.激素对盾叶薯蓣愈伤组织细胞生长和薯蓣皂苷元合成的调控.中国生物工程杂志,2002,22(3):71~84.
    72.谢彩侠,高山林,秦慧贞,等.盾叶薯蓣同源四倍体的诱导和鉴定.药物生物技术.2005,12(1):15~18,35.
    73.刘选明,李光明,秦玉芝,等.药用薯蓣高皂素新种质选育的研究.湖南农业大学学报,2004,31(3):19~22.
    74.朱延钧,树立华,张国才.武当山盾叶薯蓣生态环境和分布规律研究.资源开发与市场.1998,14(3):124~125,142。
    75.怀志萍,丁志遵,贺善安,等.盾叶薯蓣薯蓣皂苷元含量与气候因素的相关性研究.药学学报,19891,24(9):702~706.
    76.郭水良,刘鹏,王丰,吕洪飞.中国薯蓣属分布式样的模糊图论分析.浙江师大学报(自然科学版),1993,16(3):69~76.
    77.万金荣,丁志遵,秦慧贞.薯蓣科植物地理学的研究.西北植物学报,1994,14(2):128~135.
    78.张敏华,李鸿钧.湖北薯蓣属植物资源.植物资源与环境,1995,4(1):19~22.
    79.袁晓颖,祖元刚,于景华.野生盾叶薯蓣资源储量精度估算.植物研究,2003,23(1):103~105.
    80. P. K. Soderholm. Yield Trials of Steroid Producing Dioscorea on Florida's Everglades Peat Soils. Economic Botanyt. 82.
    81.丁志遵,胡同钰,王年鹤,等.土壤因素对小花盾叶薯蓣和盾叶薯蓣根茎中薯蓣皂苷元含量的影响A..南京中山植物园论文集.南京:江苏科技出版社,1983.133~134.
    82. Hegde, D. M. Comparative performance of F. B.(?)-1 and Arka Upkar cultivars of medical yam in relation to nitrogen fertilization. Indian Drgs, 1985, 22(4): 181~183.
    83. Sarma, et al. Studies on the yield of tuber and diosgenin of Dioscorea composita Hernsl. and D. foloribunda Mart. et Gal. Under various population densities. Indian Drgs, 1986, 23(6): 330~334.
    84.杨清,郑惠兰,罗明贤.菊叶薯蓣的发展前景.云南热作科技,1995,18(1):32~34.
    85.周雪林,郭可跃,朱元方,等.盾叶薯蓣引种栽培研究.中草药,1989,20(4):35~37.
    86.陈森光,王日照,周振民,等.盾叶薯蓣试管苗种植施肥初探.中国种业,2003,4:31~32.
    87.谭远友,余展深,齐迎春,等.栽培盾叶薯蓣中皂甙元含量与质量的动态变化.湖北民族学院学报,2000,18(1):17~18.
    88.蒋朝晖,麻秀萍,杨玉成,等.贵州栽培薯蓣种薯蓣皂苷元的含量变化.中药材,2001,24(8):555~556.
    89.刘铁成.穿龙薯蓣的资源调查与人工栽培.中药通讯,1985,10(12):534~535.
    90.刘铁成,严敏如.穿龙薯蓣不同生育期根茎的增长动态及薯蓣皂甙元含量变化的观察.药学通讯,1981,16(2):3~5.
    91.周瑾,陈克明,陈建忠,等.黄山药栽培试验研究.资源开发与保护,1993,9(2):83~86.
    92.徐成基,周谨,豆永泽,等.激素药源植物—盾叶薯蓣的栽培研究冲药通报,1983,8(4):3.
    93.曹玉芳,王太霞,胡正海.盾叶薯蓣营养器官薯蓣皂甙元含量的动态变化.实验生物学报,2004,37(3):221~225.
    94.李伯刚,陈建中.不同肥料对黄山药根茎产量和薯蓣皂甙元含量的影响.天然产物研究与开发,2001,13(4):27~29.
    95.王建安,徐增莱,吴国荣,等.氮磷钾对盾叶薯蓣产量及薯蓣皂苷元含量的影响.中药材,2004,27(12):891~893.
    96.李子辉.黄姜根茎折干率与皂甙元含量的相关性研究.中药材,2004,27(7):475~476.
    97.李向民,李军超,陈刚,等.盾叶薯蓣元素组成特点及其与土壤营养元素的关系研究.西北植物学报,2005,25(3):531~535.
    98.王志安,王日照,李建伟,等.施肥水平和种植密度对盾叶薯蓣试管苗亩产量和薯蓣皂甘元含量的影响.中国中药杂志,2002,27(4):300.
    99.赵庆云,谢庆华,寸湘琴,等.云南高原盾叶薯蓣低产因子分析.作物杂志,2003,(1).
    100.康阿龙,孙文基,朱朝德,等.盾叶薯蓣引种栽培调查报告.中药材,2002,25(7):465.
    101.李子辉,程新奇,万海清.盾叶薯蓣高产的关键制约因素分析.中国中药杂志,2001,26(3):203~204.
    102.齐迎春,胡诚,谭远友,等.盾叶薯蓣的栽培技术研究.氨基酸和生物资源,2003,25(1):42~43.
    103.倪喜云,张德轮,杨怀钦,等.盾叶薯蓣一年生栽培技术.中国农技推广,2005,1:34~35.
    104.杨爱民.黄姜(盾叶薯蓣)高产栽培技术.湖北农业科学,2002,6:99.
    105.朱杰英,王菊凤.盾叶薯蓣主要营养元素含量及其动态的研究.吉首大学学报,2000,21(3):7~10.
    106.朱杰英,王菊凤,陈功锡,等.野外条件下光强对盾叶薯蓣影响的初步研究.生命科学研究,2001,6(2):155~159.
    107.张友德,张君芝,秦天才,等.黄姜有性繁殖研究.华中农业大学学报,1999,18(1):8~11.
    108.秦天才,张友德,张君芝.湖北黄姜生产中的问题与对策.长江流域资源与环境,1997,6(1):35~37.
    109.张学荣,李向民.盾叶薯蓣种苗培育技术.陕西林业科技,2001(4):73~76.
    110.彭凤梅,张友发,赵庆云,等.盾叶薯蓣实生种子萌发及实生苗的生长.中国种业,2002(11):15~16.
    111.卢向阳,田云,张小明,等.几种试剂处理对黄姜种子萌发的影响.特产研究,2002,24(1):9~12.
    112.杭悦宇,黄春洪,穆森,等.盾叶薯蓣叶片形态多样性研究.云南植物研究,2004,26(4):398~404.
    113.黄春洪,杭悦宇,周义锋,等.我国盾叶薯蓣居群遗传结构分析.云南植物研究,2003,256.:641~647.
    114.郑晓琴,梁国鲁,李晓林.盾叶薯蓣自然变异类型间的比较研究.热带亚热带植物学报,2003,11(3):267~270.
    115.黄春洪,杭悦字,周义锋,等.中国盾叶薯蓣主要居群种质分析.林产化学与工业,2003. 23(2):69~73.
    116.罗跃龙,周日宝,陈茜,等.湖南省盾叶薯蓣(黄姜)种植的概况分析.湖南中医学院学报,2003,23(6):59~60.
    117.李朝阳,杨朝霞,陈功锡,等.不同居群盾叶薯蓣的生长、叶形态及皂苷元含量的研究.生命科学研究,2003,7(1):58~64.
    118.杭悦宇,夏冰,周义峰,等.武当山地区盾叶薯蓣薯蓣皂苷元含量及生物特性的相关性研究.林产化学与工业,24(2):68-72.
    119.赵岩,肖培根.我国薯蓣属甾体激素原料植物的种质资源.作物品种资源,1989,1:23~24.
    120.刘鹏,郭水良.浙江省薯蓣资源的开发利用.植物学通报,1994,11(增刊):63~65.
    121.阮运昌.激素药源植物—菊叶薯蓣中薯蓣皂甙元的质量考察.华西药学杂志,1994,9(1):205~207.
    122.秦松云,丁季春,舒抒,等.中国盾叶薯蓣资源现状及保护对策.资源开发与市场,2004,20(4):263~265.
    123.李家儒,陈克成.湖北省黄姜产业发展现状与存在问题.GAP研究与实践,2002,2(3):61~62.
    124.王泽斌.湖南省薯蓣属植物资源研究初报.中国野生植物资源,21(1):31~32.
    125.曹玉芳,林如,胡正海.盾叶薯蓣根状茎的发育解剖学研究.西北植物学报;2003,23(2):297~303.
    126.曹玉芳,胡正海.盾叶薯蓣实生苗根状茎的形态发生及薯蓣皂甙积累的研究.西北植物学报,2003,23(7):1154~1162.
    127.曹玉芳,林如,胡正海.盾叶薯蓣根状茎的发育解剖学和组织化学研究.武汉植物学研究,2003,21(4):288~294.
    128.曹玉芳,王太霞,胡正海.盾叶薯蓣根状茎不同部位和不同生长期薯蓣皂苷元含最的差异性研究.中草药,2004,35(5):562~565.
    129.郭永兵,张友德.盾叶薯蓣根茎中薯蓣皂苷元的分布.湖北农业科学,2002(5):106~107.
    130.付艳华,杜晓英,王鹏程,等.盾叶薯蓣的生长发育研究.湖北农业科学,2002,(6):98~101.
    131.唐世蓉,姜志东.盾叶薯蓣地上部分的三个新甾体皂苷.云南植物研究,1987,9(2):233~238.
    132.唐世蓉,吴余芬,庞自洁。薯蓣科植物甾体皂苷元的含量和鉴定.植物学报,1979,21(2):171~176.
    133.达世禄,唐祥怡,达晖宁,等.盾叶薯蓣中薯蓣皂苷元的反相高效液相色谱测定.色谱,1992,10(2):98~99.
    134.都述虎,王晓华,夏重道,等.RP-PHLC法测定穿龙薯蓣总皂甙中薯蓣皂甙元的含量.中国药科大学学报,2001,32(1):37.
    135.杨文远,熊楚明,张振秋.反相高效液相色谱法测定中药中薯蓣皂苷元.分析试验室,2002,21(1):74.
    136.顾生明,唐世蓉,王翔燕.薯蓣皂苷元的气象色谱分析.植物学报,1980,22(2):204-206.
    137.陶莉,达晖宁,达世禄.盾叶薯蓣中薯蓣皂苷元的气象色谱测定.中草药,1991,22(6):252~253.
    138.江天生.旋光法测定薯蓣皂甙元含量.吉首大学学报,1991,18(2):63~64.
    139.陈战国,耿征,刘谦光,等.薯蓣皂甙元的分光光度法测定.分析化学,1996,24(2):227~229.
    140.刘光东,李生德,胡军福,等.薯蓣皂苷元的分光光度法测定.郧阳师范高,等专科学校学报,2001,21(3):60.
    141.单文.用双波长薄层色谱扫描仪测定盾叶薯蓣皂甙元的含量.中草药,1981,12(1):15~16.
    142.康阿龙,孙文基,汤迎爽,等.薄层扫描法测定盾叶薯蓣中盾叶新苷和薯蓣皂苷的含量.药物分析杂志,2003,23:59~60.
    143.徐礼桑.库仑滴定法在中草药成分测定中的应用.中草药,1991,22(12):555~558.
    144.嫂纯,陆阳.吸附树脂一库仑滴定法测定盾叶薯蓣中总皂:旨的含量.中草药,62004,35(11):1309~1310.
    145. Taylor W G, Elder J L, Chang P R and Richards K W.Microdetermination of diosgenin from fenugreek (Trigonella foenum-graecum) seeds. J Agric Food Chem, 2000, 48: 5206-5210.
    146.四川省生物研究所薯蓣综合利用组.预发酵提高薯蓣皂素及淀粉综合利用的研究.植物学报,1975,17(3):242~246.
    147.吴大椿.一种发生于盾叶薯蓣上的病毒病.中草药,2000,31(11):865~866.
    148.吴大椿,严清平.盾叶薯蓣灰霉病(Botrtis cinerea)的研究.植物保护,2003,29(5):8~30.
    149.吴大椿,刘太武.田头菇危害黄姜及防治研究.湖北植保,1999,2(2):26~27.
    150.吴大椿,袁善奎,任定国,等.盾叶薯蓣茎腐病病原鉴定.湖北农学院学报,1998,18(3):224~228.
    151.吴大椿,杨海清.盾叶薯蓣除草剂的筛选.农药,2000,39(11):37.
    152.李子辉.盾叶薯蓣栽培中的草害及化控技术.植物保护,2002,5(4):42.
    153.冉西京,张忠民,朱陵侠,等.黄姜田间杂草化学防除试验初报.2001,17(1):52~53.
    154.杜晓英,付艳华,齐迎春,等.栽培盾叶薯蓣的病虫草害研究初报.氨基酸和生物资源,2003,25(1):37~39.
    155.李鹄鸣.盾叶薯蓣适光生理生态及机理研究.博士论文,2001.中南林学院.
    156.刘俊,刘选明,秦玉芝,等.Ri T-DNA对盾叶薯蓣的遗传转化及薯蓣皂甙元产生的影响.天然产物研究与开发,2005,17(1):59~64.
    157. Preston W H JR, and Haun J R. Factors involved in the vegetative propagation of Dioscorea spiculifora Hcmsl fromVincs. Proc Ame Soc Hort Sc, 1962, 80: 417-427.
    158. Prcston W H JR ctal. Several aspects of growth development and sapoganin yield of tubers of Dioscorca spiculiflora. Econ Bot, 1964, 18: 323~328.
    159. Preston W H JR. and Haun J R. Relationships of temperature and photoperiod to growth, flowering,senescence and dormancy ofDioscorea spiculifiora. Botanical gazette, 1963, 346~353.
    160. Viana A Met al. Diosgonin in leaves of Dioscorea composita. Revta brasil Bot, 1987, 10: 33o36.
    161. Passam H C. Wound repair in yam tubers. Physiological processes during repair. New Phytol. 1976, 77(2): 325~331.
    162.涂三思,秦天才.高温胁迫对黄姜叶片脯氨酸、可溶性糖和丙二醛含量的影响.湖北农业科学,2004,(4):98~100.
    163.肖春桥,李家儒,朱广慧.多效唑对盾叶薯蓣叶片活性氧代谢与膜脂过氧化反应的影响.武汉植物学研究,2003,21(2):170~174.
    164.陈世忠.石墨炉原子吸收光谱法测定中药黄姜中微量镉的研究光谱学与光谱分析,2003,23(5):993~994.
    165.陈灵芝,马克平.生物多样性科学:原理与实践.上海:上海科学技术出版社,2001.93.
    166.施立明,贾旭,胡志昂.遗传多样性.见:陈灵芝.中国的生物多样性.北京:科学出版社,1993.99.
    167.葛颂.保护生物学(蒋志刚,马克平)—遗传多样性.杭州:浙江科学技术出版社,1997,11.
    168. Bruno Baur, Schmid B. Spatial and temporal patterns of genetic diversity within species. In: Kevin J. Gaston ed. Biodiversity: A Biology of Numbers and Difference. London: Blackwell Science Institute, 1996. 169.
    169.Wodd Resourcea Inatitute(WRI),等.中国科学院生物多样性委员会译.全球生物多样性策略北京:中国标准出版社,1993.
    170.刘祖洞.遗传学(第二版).北京:高等教育出版社,1991.
    171. Nei M. Molecular Evolution Genetics. New York: Columbia University Press, 1987.
    172. Nei M, W. H Li. Mathematical Model for Studying Genetic Variation in terms of Restriction Endonnuclese. Proc. Natl. Acad. Sci. U.S.A. 1979, 76: 5269.
    173. Lynch M, Crease T J. The analysis of population surveyed data on DNA sequence variation. Mol. Biol. Evol. 1990, 7: 377.
    174. Miyamota and Cracraft. Phylogenetic Analysis of DNA Sgue Flces. In: Hillis, Moritz and Mable. Molecular Sysmatics. 1996.
    175. Chen Xiaofeng, Shipeng Li. Estimation of nucleotide diversity from RAPD date. In: N Li & Y Chen ed. Proceedings of International Conferece on Animal Biotechnology. Beijing: International Academic Publishers. 1997, 185.
    176. Williams J, Kubelik A, Livak, et al. DNA polymprphisms amplified by arbitrary primers are useful as genetic markers. NUCL. Acids. Res, 1990,18:6531~6535.
    177. Welsh J, M: AcClelland. Fingerprinting genomes using PCR with arbitrary palmers. Acids. Res, 1990,18:7213~7218.
    178.汪小全,周喻萍,张大明,等.RAPD应用于遗传多样性和系统学研究中的问题.植物学报,1996,38(12):954~962.
    179. Parker P G, Snow A A, Schug M D, et al. What molecular can tell us about populations: chooseing and using a molecular marker. Ecology,1998,79 (2):361.
    180.郭军,许勇,寿森炎,等.西瓜种质资源遗传关系的RAPD分析.植物遗传资源科学.2002,3(1):7-13.
    181.李云海,钱前,曾大力,等.我国主要杂交水稻亲本的RAPD鉴定及遗传关系研究.作物学报,2000,26(2):171~176.
    182. Thorman C E,Ferreira M E, Camargo L E Aet al.Comparison of RFLP and RAPD markers to estimating genetic relatianships within and among crucifereus species.Theor Appl Genet.1994,80:973~980.
    183.陈云鹏,曹家树,缪颖,等.芸苔类蔬菜基因组DNA遗传多样性RAPD分析.浙江大学学报,2001.26(2):131~136.
    184. Wilkia S E. Isaac P G. Slater R J. Random amplified polymorphic DNA RAPD markers for genetic analysis in Allum.Theor Appl Genet.1993,06:497~504.
    185.栾时雨,苏乔,李海涛,等.利用RAPD技术快速鉴定番茄杂种纯度.园艺学报,1998,(3):247~252.
    186.庄木,王晓武,杨丽梅,等.利用RAPD方法鉴定两个番茄品种的纯度.中国蔬菜,1999,(5):8~9.
    187.王晓武,方智远,孙培田,等.一个与甘蓝显性雄性不育基因连锁的RAPD标记.园艺学报,1998,(2);197~199.
    188.曹家树,曹寿椿,易清明.白菜及其相邻类群基因组DNA和RAPD分析.园艺学报,1995,22(1):47~52.
    189.漆小泉,朱德蔚,沈镝,等.大白菜和紫菜苔自交系染色体组DNA的RAPD分析.园艺学报,1995,22(3):256~262.
    190.乔爱民,刘佩瑛,雷建军,等.利用RAPD标记鉴定芥菜(Brassica juncea Coss.)品种.中山大学学报,1998,37(2):73~76.
    191.张海英,王永健,许勇,等.黄瓜种质资源遗传亲缘关系的RAPD分析.园艺学报,1998,(4):345~350.
    192.庄飞云,庄飞云,陈劲枫.黄瓜栽培种、近缘野生种、种间杂种及其后代的RAPD分析.园艺学报,2003,30(1):47~50.
    193.张海英,王永健,许勇,等.黄瓜种质资源遗传亲缘关系的RAPD分析.园艺学报,1998,(4):345~350.
    194.许东河,高忠,盖均镒,等.中国野生大豆与栽培大豆等位基因RFLP和RAPD标记的遗传多样性与演化趋势分析.中国农业科学,1999,32(6):16-22.
    195.胡志昂,张亚平.中国动植物的遗传多样性.杭州:浙江科技出版社,1997.
    196.韩春艳,孙卫邦,高连明.濒危植物三棱栎遗传多样性的RAPD分析.云南植物研究,2004,265:513~518.
    197.恽锐,钟敏,王洪新,等.北京东灵山辽东栎群体DNA多样性的研究.植物学报,1998,40(2): 169-175.
    198.恽锐,王洪新,钟敏,胡志昂,魏伟,钱迎倩.蒙古栎、辽东栎的遗传分化:从形态到DNA.植物学报,1998,40(11):1040~1046.
    199.魏伟,王洪新,胡志昂,等.毛乌素沙地柠条群体分子生态学初步研究:RAPD证据.生态学报,1999,19(1):16~22.
    200.裴颜龙,邹喻苹,尹蓁,等.矮牡丹与紫斑牡丹RADP分析初报.植物分类学报.1995,33(4) 350~356.
    201.汪小全,刘正字.银杉遗传多样性的RADP分析.中国科学(C辑),1996,6(5):436~441.
    202.李宽钰,杨自湘.青杨的遗传分化.植物学报,1997,39(8):753~758.
    203.施苏华,唐绍清,陈月琴,等.11种金花茶植物的RAPD分析及其系统学意义.植物分类学报,1998.36(4):317~322.
    204.朱晓琴,贺善安.苍术种内遗传多样性分析.植物资源与环境,1995,4(2):1~6.
    205.梁风山,罗耀武.多倍体及其在农业生产中的应用.国外农学—杂粮作物,1999,19(2):20~23.
    206.纪风高,邓景扬.秋水仙素与二甲基亚枫混合水溶液处理小麦.黑麦杂交一代苗进行染色体加倍的试验.遗传,1987,9(3):1~4,10.
    207.乔传卓,吴美枢,戴富宝,等.菘蓝多倍体育种的研究.植物学报,1989,31(9):678.
    208.朱英,来平凡.石菖蒲的研究近况.浙江中医学院学报,2001,25(3):80.
    209.罗跃龙,彭菲,刘军.杭白芷的多倍体诱导与培育.中国中药杂志,2004,29(2):186.
    210.吕世民,梁可钧,葛传吉,等.怀牛膝多倍体育种的研究.中药通报,1988,13(7):395.
    211.运泓,杨治琴,张治国.化学诱变同源多倍体当归育种初报.中药材,1980,11(1):9.
    212.张俊莲,张志蒙.秋水仙素处理当归愈伤组织分化及植株生长发育的影响.甘肃农业科技,1999,2:14.
    213.陈绍潘,陈绍裘,杨英华,等.秋水仙碱诱变甜菊多倍体的研究.武汉植物学研究,1995,13(1):1.
    214.陈柏君,高山林,卞云云.黄芩组织培养同源四倍体的诱导.植物资源与环境学报,2000,90:9.
    215.乔传卓,崔熙.药用植物多倍体的应用.中药材科技,1981,4(4):40.
    216.艾建国,高山林.丹参同源四倍体的诱导、鉴定及有效成分含量分析.药用生物技术,2003,10(6):372~376.
    217.李竟雄,宋同明.植物细胞遗传学.科学出版社,1997.
    218.孙宗修,卓丽圣,程式华,等.水稻花培技术的改进及其在杂交水稻育种中的应用.农业生物技术学报,1997,3:244~251.
    219.颜昌敬.农作物组织培养.上海:科学技术出版社,1991,283~289.
    220.胡道芬.植物花药育种进展.北京:中国农业科技出版社,1995,6~11.
    221.李娈,蒋道松,刘硕.单子叶植物花药培养的研究进展.湖南农业大学学报.2005 11,31(2):41~46.
    222.马克平.生物多样性及其研究进展.中国生物多样性国情研究报告.北京:中国环境出版社,1998
    223. UNEP. Global Biodiversity Assessment. 1992.
    224.张成合,宋长志,王淑芳.多倍体育种综述.河北农业大学学报,1988,11(2):136~139.
    225.李枸.染色体遗传导论.湖南科学出版社,1991.
    226.周朴华,何立珍,刘选明.组织培养中用秋水仙素诱发黄花菜同源四倍体的研究.中国农业科学,1995.28(1):49~55.
    227.薛焱.不同居群及株系盾叶薯蓣的形态生理特性研究.硕士论文,2005,5.
    228. Murry HG, Thomspon WF. Rapid isolation of weight DNA. Nucleic Acid Res, 1980, 8: 4312~4322..
    229.薛焱,秦天才,张友德.不同薯蓣皂苷元的盾叶薯蓣遗传关系的RAPD分析.中草药,2006,37 (1):115~118.
    230.李晶,秦天才.盾叶薯蓣居群亲缘关系的RAPD分析.湖北农业科学.2005,4:20~22.
    231. Peter Schols, Carol A. Furness, Paul Wilkin, et al. Pollen morphology of Dioscorea (Dioscoreaceae) and its relation to systematics. Bot. Jou. of the Linnean Society, 2003,143, 375~390.
    232. Galitski T, Saldanha A J, Styles C A, et al. Ploidy regulation of gene expression. Science, 1999,285:251~254.
    233. Yang J(杨继). The formation and evolution of polyploidy genomes in plants.Acts Phytotaxon Sin(植物分类学报),2001,39(4):357~371.
    234.常青,周开亚.分子进化研究中系统发生树的重建.生物多样性,19986(1):55~62.
    235. Felsenstein J. Phylogenies from molecular sequences: Inference and reliability. A nnu. Rev. Genet, 1988,22:521~565.
    236.胡志昂,张亚平.中国动植物的遗传多样性.杭州:浙江科学技术出版社,1997.
    237. Felsenstein J. Phylogeny Inference Package (PHYL IP), Version 3. 5. University of Washington, Seattle. 1993.
    238. Swofford D L. Phylogenetic Analysis Using Parsimony (PAUP), Version 3. 1.1. University of Illinois, Champaign. 1993.
    239. Kumar S,Tamura K, Nei M. MEGA: Molecular Evolutionary Genetics Analysis, V.ersion 1. 01. The Pennsylvania State University, University Park, PA 16802. 1993.
    240. Excoffier L. AMOVA: Analysis of Molecular Variance, Version 1. 55. Genetics and Biometry Lab, University of Geneva. 1993.
    241. Armstrong J, Gibbs A,Peakall R et al. RAPD istance Programs; Version 1. 04 for the analysis of patterns of RAPD fragments. Canerra: Australian National University. 1996.
    242. Demeke T, Adams R P. Chibbar R. Potential taxonomic use uf random amplified polymorphic DNA (RAPD) A case study in Brassica. Theor Appl. Genet,1992,84:990~994.
    243. Hatward T. Stalker T. Larue E et al. Use of single-primer DNA amphficatiuns in genetic studies of peanut (Arachis hypoguea L.).Plant Mol Biol. 1992,10: 315~325.
    244. Wilkia S E. Isaac P G.Slater R J. Random amplified polymorphic DNA RAPD markers for genetic analysis in Allum.Theor Appl Genet.1993,06:497~504.
    245.彭尽晖,张良波,彭晓英.秋水仙素在植物倍性育种中的应用进展.湖南林业科技,2004,31(5):22~25
    246.任清,罗耀武,柳术杰.人工诱变四倍体玫瑰香葡萄的遗传稳定性研究.园艺学报,2000,27(4):285~286.
    247.武振华,牛炳韬,王新宇.药用植物染色体加倍的研究进展.西北植物学报,2005,25(12):2569~2571.
    248.彭卫欣,谢小亮.药用植物多倍体研究进展.河北农业科学,2002,6(3):47~50.
    249.丁世萍,严菊强,季道藩.糖类在植物组织培养中的效应.植物学通报,1998,15(6):42~46.
    250.张宝红,刘方,姚长兵,等.糖源对棉花花药培养的影响。江西农业大学学报,1998,20(3):311~316.
    251.李胜,杨德龙,武季玲,等.扁桃花药培养初报.甘肃农业大学学报,2005,40(5):617~620.
    252.贾勇炯,陈放.在烟草花药培养中诱导胚状体发生的研究.四川大学学报,1996,33(5):605~608.
    253.柯卫东,彭静,刘玉平,等.试管藕诱导技术研究,武汉植物学研究.2001,19(2):173-175.
    254.中国医学科学院药物研究所.中草药现代研究.北京:北京医科大学中国协和医科大学联合出版社.1996:226~254.
    255.张秀清,王志武,刘玉敬,等.蝴蝶兰实生苗不同器官的离体培养.植物学通报.1996,13(1):50~52
    256.周俊辉,叶超宏,陈旭高.蝴蝶兰原球茎增殖培养的研究.植物生理学通讯.2002,15(3):13~17.
    257. Omidiji O, Okpuzor J. Time course of PPO-related browning of yams. Journal of the Science of Food and Agriculture.1996,70(2):190~196.
    258.符策,陆祖正,周婧,等.盾叶薯蓣开花结果调查.广西热带农业,2005(1):10~12.
    259.赵庆云,寸湘琴,杨燕,等.云南高原几个盾叶薯预种质特征特性及栽培要点.种子,2003(1):41~42.
    260.周媛,胡春根,时光,姚家玲.四倍体盾叶薯蓣生物学特性的研究.武汉植物学研究,2005,23(4):358~362.
    261.何立珍,刘选明,周朴华.黄花菜HAC—大花长嘴子花同源四倍体选育.湖南农业科学,1993(4):23~24.
    262.黄济明.百合组织培养和试管内诱发多倍体试验.园艺学报,1983(2):125~127.
    263.郑思乡,邬明芳,李宗道,等.不同倍性苎麻同工酶及光合作用研究.中国麻作,1999,21(2):1~4.
    264.许大全.光合速率、光合速率和作物产量.生物学通报,1999,34(8):8~10.
    265.罗勇军,罗光富,黄应平,等.过氧化物酶模拟及应用研究进展.分析测试学报,2004,23(5):136~142.
    266.孟学平,杨恒山,郭宏,等.盐胁迫对冬小麦过氧化物酶同工酶的影响.华北农学报,2000,15(4):62~67.
    267.詹嘉红,蓝宗辉.外源ABA对低温胁年头水稻过氧化物酶同工酶的影响.生物技术,2003,13(1):7~9.
    268. Walker J R, Ferrar P H. Diphenol oxidase, enzyme-catalysed browning and plant disease resistance. Biotechn Gen Eng Rev, 1998,15:457~498.
    269. Richard F F, Gauillard F A. Oxidation of chlorogenic acid, catechins,and 4-methylcatechol in model solutions by combinations of pear(Pyrus communis cv.Willians) polyphenol oxidase and peroxidase:a possible involvement of peroxidase in enzymatic browning. Agri Food Chem,1997,45:2472~2476.
    270. Nicolas J J, Richard~Forget F, Goupy P, et al. Enzymatic browning reactions in apple and apple products. Crit Rev Food Sci Nutur, 1994,34:109-157.
    271. Lepoz Serrano M, Barcelo A R. Purification and characterization of a basic peroxidase isoenzyme from strawberry. Food Chem,1996,55:133~137.
    272.蒋选利,李振岐,康振生.过氧化物酶与植物抗病性研究进展.西北农林科技大学学报,2001,29(6):124~129.
    273. Liu D L, Yu B C, Sun Y Y. Effects of the temperature stress on SOD and CAT activities and their isorymes of rice seedling. J Central China Normal Uniu: Nat Sri, 1994, 28(4): 525~528.
    274.朱松玲,吕广宇.空气污染对几种植物过氧化氢酶活性的影响.泰山学院学报,2003,25(6):88~90.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700