苹果多倍体种质创新及鉴定评价研究
|
摘要
苹果是世界性果树,世界上相当多的国家都将其列为主要消费果品。由于果树多倍体一般具有生长旺盛、果实大且少籽或无籽、产量高、适应性和抗逆性强等特点,且能够利用无性繁殖的方式固定其优良性状,使之保持稳定而不分离,所以多倍体育种一直被认为是创造苹果新品种的重要途径。本研究以金冠、嘎拉、富士苹果的成熟胚和寒富、烟嘎1号的离体叶片为试材,通过秋水仙素诱变技术和二倍体与多倍体杂交的方法,获得苹果多倍体新种质。同时在分子水平上探究苹果四倍体与二倍体之间的差异,为阐明苹果多倍体基因组进化机理奠定基础。主要研究结果如下:
1.建立了苹果成熟胚离体诱导四倍体的技术体系。金冠的成熟胚在0.5%的秋水仙素+1%二甲基亚砜的混合溶液中浸泡48h后接种在MS+TDZ 1.0 mg·L~(-1)+IBA 0.5 mg·L~(-1)培养基上进行再生培养,诱变率达10.6%。
2.建立了以寒富、烟嘎苹果的试管苗叶片离体诱导四倍体的技术体系。寒富苹果叶片在附加15、30、60、120 mg·L~(-1)秋水仙素的液体再生培养基(MS+1.0 mg·L~(-1) TDZ+0.5 mg·L~(-1) NAA+200 mg·L~(-1) LH)中静置暗培养5 d,各个浓度处理均诱导出四倍体植株,诱变率在5.3%～22.2%之间。寒富、烟嘎1号苹果叶片在附加50 mg·L~(-1)秋水仙碱固体再生培养基上处理5 d亦获得了四倍体植株,诱变率分别为20.0%和15.0%。
3.将诱变获得的倍性嵌合体植株的叶片接种到MS+TDZ 1.0 mg·L~(-1)+IBA 0.5 mg·L~(-1)+蔗糖30 g·L~(-1)+琼脂6 g L~(-1)培养基上进行再生,从金冠、嘎拉、富士的倍性嵌合体中均分离出了同质突变体。
4.比较了3种获得苹果三倍体的方案:(1)通过四倍体与二倍体杂交获得三倍体;(2)通过三倍体与二倍体杂交获得三倍体;(3)在三倍体实生后代中筛选三倍体。表明最好的方法是二倍体与四倍体间的杂交,出现三倍体比率明显高于其他两种方法。
5.苹果多倍体植株生长健壮,枝条粗壮,叶片大而圆,粗糙,叶厚,叶边锯齿较多,节间较短。多倍体的气孔比二倍体和非整倍体的气孔大得多,在气孔密度上,多倍体小于二倍体和非整倍体。苹果三倍体实生后代中不同倍性植株在形态上差异显著,根据植株形态和叶形指数,可以容易地将多倍体(三倍体、四倍体)植株与二倍体植株和非整倍体植株区分开来。
6.二倍体苹果品种富士和烟嘎1号叶绿素a、叶绿素b、叶绿素总含量和类胡萝卜素分别低于其同源四倍体天星和四倍体烟嘎1号品种,但叶绿素a/b比值与倍性没有表现出一致的变化规律。对二倍体品种富士和烟嘎1号及其各自同源四倍体品种的叶绿素荧光参数日变化规律进行了测定,结果显示二倍体品种富士和烟嘎1号Fo日平均值分别为214.6和208.0,分别较天星和四倍体烟嘎1号高出9.4和8.3。二倍体品种富士和烟嘎1号的PI值分别小于其同源四倍体品种天星和四倍体烟嘎1号。
7.通过电镜观察,二、四倍体苹果花粉的大部分侧面观均为长椭圆形,极面观为三角形,具3纵沟孔;花粉表面为纵向条状纹饰,有穿孔。四倍体花粉量明显比二倍体少,非椭圆形花粉粒特别多。四倍体P/E值为1.8,二倍体为2.0,差异显著。四倍体胚囊存在不同程度的异常结构。
8.采用ISSR分子标记技术对富士、烟嘎1号、寒富的二倍体及其同源四倍体进行PCR扩增。8条引物共扩增出65条180～2500bp谱带,其中24条带具有多态性,占36.9%,每个引物可扩增出0～5条多态性带,平均为3条。三组苹果(富士和天星、寒富和四倍体寒富、烟嘎1号和四倍体烟嘎1号)之间有所差异,但富士、烟嘎1号、寒富的二倍体及其同源四倍体间的遗传相似系数全部为1,不能区分。
9.利用高效液相色谱仪测定了苹果二倍体品种及其同源四倍体叶片DNA水解样中胞嘧啶和5-甲基胞嘧啶的含量。四倍体天星与二倍体富士相比,天星的DNA甲基化程度降低,为29.33%,富士为35.34%。四倍体寒富和二倍体寒富甲基化程度分别是14.37%和14.49%,差异不大。
10.本研究共获得金冠、嘎拉、富士成熟胚诱导的同质四倍体251株,寒富叶片诱导的四倍体20株,烟嘎1号叶片诱导的四倍体13株。从四倍体与二倍体杂交后代、三倍体与二倍体杂交后代及三倍体实生后代中共筛选出三倍体46株、四倍体5株。上述多倍体种质的获得为选育苹果多倍体品种奠定了重要基础。
The apple is the world fruit,and most countries in the world rank it with the main consuming fruits. Because polyploidy fruit trees generally have the characteristics of vigorous growing,big fruit size, fewer seeds or seedless,high production,strong adaptability and resistance,and could keep excellent characteristics with asexual propagation,polyploidy breeding is considered as the importance approach of create new apple cultivars.This study obtained apple novel polyploidy germplasm with the mature embryos of 'Golden Delicious','Gala','Fuji' and the in vitro leaf of 'Yan 1 Gala','Hanfu' as materials by colchicine induction and polypolid hybridization.Besides on that,we analyzed the difference between apple tetraploid and diploid on molecule level,which will lay a foundation for the work on elucidating the evolution mechanism of apple polyploidy.The main results are as follows:
1.The system of induction of tetraploid from apple mature embryos in vitro was successfully established.The induction rates was 10.6%when the mature embryos of 'Golden Delicious' apple were treated with 0.5%colchicines and 1%DMSO for 48h and then cultured on MS medium supplemented with TDZ 1.0mg·L~(-1),IBA 0.5 mg·L~(-1).
2.In vitro tetraploid induction of 'Hanfu' and 'Yan 1 gala' apple were studied by treating leaf with colchicines solution.The inducing percentage was from 5.3%to 22.2%when the leavess of 'Hanfu' were treated in liquid medium(MS+TDZ 1.0 mg·L~(-1)+NAA 0.5 mg·L~(-1)+LH 200 mg·L~(-1)) supplemented with colchicines 15 mg·L~(-1),30 mg·L~(-1),60 mg·L~(-1) or 120 mg·L~(-1) for five days at dark.And tetraploid plantlets were also obtained from 'Hanfu' and 'Yan 1 gala' when the leaves were treated on solid medium supplemented with colchicines for five days at dark.The induction rates were 20.0%and 15.0% for 'Hanfu' and 'Yan 1 gala' respectively.
3.The leaves of ploidy chimeras were inoculate to culture medium which contained MS+TDZ 1.0 mg·L~(-1)+IBA 0.5 mg·L~(-1)+sucrose30 g·L~(-1)+ Agar 30g·L~(-1) for adventitious shoots regeneration.The homogeneity mutants could be separated form the 'Golden delicious','Gala' and 'Fuji' ploidy chimeras.
4.Comparing the three schemes for obtaining apple triploid plants:(1) hybridization between diploid and tetraploid plant;(2) crossing between diploid and triploid plant;(3) selecting triploid plants from triploid seedling progenies.Result show that the best methed was the hybridization between diploid and tetraploid plant,as the percentage of triploid plants obviously higher than that using other schemes.
5.Apple polyploid plants were vigorous with strong shoots and short internodes,and its leaves were big,round,coarse,and thick with more teeth,.Stomas of triploid seedling progenies were bigger than that of diploid and aneuploid plant,but the density of stomas was less than that in diploid and aneuploid plant. Among the polyploid seedling progenies of apple,different ploidy plants had marked discrimination in plant shape.Based on plant shape and index of leaf shape,polyploid(tetraploid and triploid) plants could easily be distinguished from the diploid plants and the aneuploid plants.
6.The chlorophyll a,chlorophyll b,the total amount of chlorophyll and carotenoid of diploid apple cultivars 'Fuji' and 'Yan 1 Gala' were below their autotetraploid 'Tensei' and 'tetraploid Yan 1 Gala', respectively.However,chlorophyll a/b ratio have not showed consistent changes.The chlorophyll fluorescence parameters changes in the law of diploid varieties 'Fuji' and 'Yan 1 Gala' and their respective autotetraploid species were measured and the results showed that FO averages of diploid varieties 'Fuji' and 'Yan 1 Gala' were 214.6 and 208.0,which are higher than the 'Tensei' and 'tetraploid Yan 1 Gala'-9.4 and 8.3.The 'PI' on diploid varieties Fuji and 'Yan I Gala' were lower than their autotetraploid 'Tensei' and 'tetraploid Yan 1 Gala' respecitively.
7.The diploidy and tetraploidy pollens of apples were observed by electron microscope,the pollens have three longthways channels,most of their profiles were oblong and polars were triangle.The pollen surfaces have longways striations and perforations.The pollen quantity of tetraploids were less than that of diploids,and nonablong pollens were only few.P/E value of tetraploids was 1.8,and that of diploids was 2.0,which have significant difference.
8.The 'Fuji','Yan 1 Gala' and 'Hanfu' of diploid and autotetraploid were analyzed with ISSR markers.A total of 65 bands,ranged from 180 to 2500 bp,were amplified with eight primers. Twenty-four bands were polymorphic,accounting for 36.9%,and each primer can amplify from 0 to 5 polymorphic bands,with 3 bands at average.Three groups of apples('Fuji' and 'Tensei','Hanfu' and 'tetraploid Hanfu','Yan 1 Gala' and 'tetraploid Yan 1 Gala') have difference,but the genetic similarity coefficients between diploidys and its autotetraploidys of 'Fuji','Yan 1 Gala' and 'Hanfu' all were 1,and they could not distinguish each other.
9.The content of cytosine and 5-methyl cytosine in hydrolysis DNA samples extracted from leaves of the diploid of apple and its autotetraploid was identified used HPLC.The 'Tensei' had low DNA methylation which was 29.33%and 'Fuji' was 35.34%.The methylation degree of 'Hanfu' diploid and tetraploid were 14.37%and 14.49%respectively,with no significant difference.
10.Two hundred and fifty-one homogeneity tetraploid trees were induced from embryos of apple cultivars 'Golden Delicious','Gala' and 'Fuji'.Twenty and thirteen tetraploid plants were induced from leaves of 'Hanfu' and 'Yan Gala 1' respectively.Forty-six triploid tress and 5 teraploid trees were obtained from the offsprings of teraploid crossing diploid,the offsprings of triploid crossing diploid,and the self-progenies of triploid.These polyploidy germplasms laid the important foundation for breeding polyploid apple cultivars.
1.建立了苹果成熟胚离体诱导四倍体的技术体系。金冠的成熟胚在0.5%的秋水仙素+1%二甲基亚砜的混合溶液中浸泡48h后接种在MS+TDZ 1.0 mg·L~(-1)+IBA 0.5 mg·L~(-1)培养基上进行再生培养,诱变率达10.6%。
2.建立了以寒富、烟嘎苹果的试管苗叶片离体诱导四倍体的技术体系。寒富苹果叶片在附加15、30、60、120 mg·L~(-1)秋水仙素的液体再生培养基(MS+1.0 mg·L~(-1) TDZ+0.5 mg·L~(-1) NAA+200 mg·L~(-1) LH)中静置暗培养5 d,各个浓度处理均诱导出四倍体植株,诱变率在5.3%～22.2%之间。寒富、烟嘎1号苹果叶片在附加50 mg·L~(-1)秋水仙碱固体再生培养基上处理5 d亦获得了四倍体植株,诱变率分别为20.0%和15.0%。
3.将诱变获得的倍性嵌合体植株的叶片接种到MS+TDZ 1.0 mg·L~(-1)+IBA 0.5 mg·L~(-1)+蔗糖30 g·L~(-1)+琼脂6 g L~(-1)培养基上进行再生,从金冠、嘎拉、富士的倍性嵌合体中均分离出了同质突变体。
4.比较了3种获得苹果三倍体的方案:(1)通过四倍体与二倍体杂交获得三倍体;(2)通过三倍体与二倍体杂交获得三倍体;(3)在三倍体实生后代中筛选三倍体。表明最好的方法是二倍体与四倍体间的杂交,出现三倍体比率明显高于其他两种方法。
5.苹果多倍体植株生长健壮,枝条粗壮,叶片大而圆,粗糙,叶厚,叶边锯齿较多,节间较短。多倍体的气孔比二倍体和非整倍体的气孔大得多,在气孔密度上,多倍体小于二倍体和非整倍体。苹果三倍体实生后代中不同倍性植株在形态上差异显著,根据植株形态和叶形指数,可以容易地将多倍体(三倍体、四倍体)植株与二倍体植株和非整倍体植株区分开来。
6.二倍体苹果品种富士和烟嘎1号叶绿素a、叶绿素b、叶绿素总含量和类胡萝卜素分别低于其同源四倍体天星和四倍体烟嘎1号品种,但叶绿素a/b比值与倍性没有表现出一致的变化规律。对二倍体品种富士和烟嘎1号及其各自同源四倍体品种的叶绿素荧光参数日变化规律进行了测定,结果显示二倍体品种富士和烟嘎1号Fo日平均值分别为214.6和208.0,分别较天星和四倍体烟嘎1号高出9.4和8.3。二倍体品种富士和烟嘎1号的PI值分别小于其同源四倍体品种天星和四倍体烟嘎1号。
7.通过电镜观察,二、四倍体苹果花粉的大部分侧面观均为长椭圆形,极面观为三角形,具3纵沟孔;花粉表面为纵向条状纹饰,有穿孔。四倍体花粉量明显比二倍体少,非椭圆形花粉粒特别多。四倍体P/E值为1.8,二倍体为2.0,差异显著。四倍体胚囊存在不同程度的异常结构。
8.采用ISSR分子标记技术对富士、烟嘎1号、寒富的二倍体及其同源四倍体进行PCR扩增。8条引物共扩增出65条180～2500bp谱带,其中24条带具有多态性,占36.9%,每个引物可扩增出0～5条多态性带,平均为3条。三组苹果(富士和天星、寒富和四倍体寒富、烟嘎1号和四倍体烟嘎1号)之间有所差异,但富士、烟嘎1号、寒富的二倍体及其同源四倍体间的遗传相似系数全部为1,不能区分。
9.利用高效液相色谱仪测定了苹果二倍体品种及其同源四倍体叶片DNA水解样中胞嘧啶和5-甲基胞嘧啶的含量。四倍体天星与二倍体富士相比,天星的DNA甲基化程度降低,为29.33%,富士为35.34%。四倍体寒富和二倍体寒富甲基化程度分别是14.37%和14.49%,差异不大。
10.本研究共获得金冠、嘎拉、富士成熟胚诱导的同质四倍体251株,寒富叶片诱导的四倍体20株,烟嘎1号叶片诱导的四倍体13株。从四倍体与二倍体杂交后代、三倍体与二倍体杂交后代及三倍体实生后代中共筛选出三倍体46株、四倍体5株。上述多倍体种质的获得为选育苹果多倍体品种奠定了重要基础。
The apple is the world fruit,and most countries in the world rank it with the main consuming fruits. Because polyploidy fruit trees generally have the characteristics of vigorous growing,big fruit size, fewer seeds or seedless,high production,strong adaptability and resistance,and could keep excellent characteristics with asexual propagation,polyploidy breeding is considered as the importance approach of create new apple cultivars.This study obtained apple novel polyploidy germplasm with the mature embryos of 'Golden Delicious','Gala','Fuji' and the in vitro leaf of 'Yan 1 Gala','Hanfu' as materials by colchicine induction and polypolid hybridization.Besides on that,we analyzed the difference between apple tetraploid and diploid on molecule level,which will lay a foundation for the work on elucidating the evolution mechanism of apple polyploidy.The main results are as follows:
1.The system of induction of tetraploid from apple mature embryos in vitro was successfully established.The induction rates was 10.6%when the mature embryos of 'Golden Delicious' apple were treated with 0.5%colchicines and 1%DMSO for 48h and then cultured on MS medium supplemented with TDZ 1.0mg·L~(-1),IBA 0.5 mg·L~(-1).
2.In vitro tetraploid induction of 'Hanfu' and 'Yan 1 gala' apple were studied by treating leaf with colchicines solution.The inducing percentage was from 5.3%to 22.2%when the leavess of 'Hanfu' were treated in liquid medium(MS+TDZ 1.0 mg·L~(-1)+NAA 0.5 mg·L~(-1)+LH 200 mg·L~(-1)) supplemented with colchicines 15 mg·L~(-1),30 mg·L~(-1),60 mg·L~(-1) or 120 mg·L~(-1) for five days at dark.And tetraploid plantlets were also obtained from 'Hanfu' and 'Yan 1 gala' when the leaves were treated on solid medium supplemented with colchicines for five days at dark.The induction rates were 20.0%and 15.0% for 'Hanfu' and 'Yan 1 gala' respectively.
3.The leaves of ploidy chimeras were inoculate to culture medium which contained MS+TDZ 1.0 mg·L~(-1)+IBA 0.5 mg·L~(-1)+sucrose30 g·L~(-1)+ Agar 30g·L~(-1) for adventitious shoots regeneration.The homogeneity mutants could be separated form the 'Golden delicious','Gala' and 'Fuji' ploidy chimeras.
4.Comparing the three schemes for obtaining apple triploid plants:(1) hybridization between diploid and tetraploid plant;(2) crossing between diploid and triploid plant;(3) selecting triploid plants from triploid seedling progenies.Result show that the best methed was the hybridization between diploid and tetraploid plant,as the percentage of triploid plants obviously higher than that using other schemes.
5.Apple polyploid plants were vigorous with strong shoots and short internodes,and its leaves were big,round,coarse,and thick with more teeth,.Stomas of triploid seedling progenies were bigger than that of diploid and aneuploid plant,but the density of stomas was less than that in diploid and aneuploid plant. Among the polyploid seedling progenies of apple,different ploidy plants had marked discrimination in plant shape.Based on plant shape and index of leaf shape,polyploid(tetraploid and triploid) plants could easily be distinguished from the diploid plants and the aneuploid plants.
6.The chlorophyll a,chlorophyll b,the total amount of chlorophyll and carotenoid of diploid apple cultivars 'Fuji' and 'Yan 1 Gala' were below their autotetraploid 'Tensei' and 'tetraploid Yan 1 Gala', respectively.However,chlorophyll a/b ratio have not showed consistent changes.The chlorophyll fluorescence parameters changes in the law of diploid varieties 'Fuji' and 'Yan 1 Gala' and their respective autotetraploid species were measured and the results showed that FO averages of diploid varieties 'Fuji' and 'Yan 1 Gala' were 214.6 and 208.0,which are higher than the 'Tensei' and 'tetraploid Yan 1 Gala'-9.4 and 8.3.The 'PI' on diploid varieties Fuji and 'Yan I Gala' were lower than their autotetraploid 'Tensei' and 'tetraploid Yan 1 Gala' respecitively.
7.The diploidy and tetraploidy pollens of apples were observed by electron microscope,the pollens have three longthways channels,most of their profiles were oblong and polars were triangle.The pollen surfaces have longways striations and perforations.The pollen quantity of tetraploids were less than that of diploids,and nonablong pollens were only few.P/E value of tetraploids was 1.8,and that of diploids was 2.0,which have significant difference.
8.The 'Fuji','Yan 1 Gala' and 'Hanfu' of diploid and autotetraploid were analyzed with ISSR markers.A total of 65 bands,ranged from 180 to 2500 bp,were amplified with eight primers. Twenty-four bands were polymorphic,accounting for 36.9%,and each primer can amplify from 0 to 5 polymorphic bands,with 3 bands at average.Three groups of apples('Fuji' and 'Tensei','Hanfu' and 'tetraploid Hanfu','Yan 1 Gala' and 'tetraploid Yan 1 Gala') have difference,but the genetic similarity coefficients between diploidys and its autotetraploidys of 'Fuji','Yan 1 Gala' and 'Hanfu' all were 1,and they could not distinguish each other.
9.The content of cytosine and 5-methyl cytosine in hydrolysis DNA samples extracted from leaves of the diploid of apple and its autotetraploid was identified used HPLC.The 'Tensei' had low DNA methylation which was 29.33%and 'Fuji' was 35.34%.The methylation degree of 'Hanfu' diploid and tetraploid were 14.37%and 14.49%respectively,with no significant difference.
10.Two hundred and fifty-one homogeneity tetraploid trees were induced from embryos of apple cultivars 'Golden Delicious','Gala' and 'Fuji'.Twenty and thirteen tetraploid plants were induced from leaves of 'Hanfu' and 'Yan Gala 1' respectively.Forty-six triploid tress and 5 teraploid trees were obtained from the offsprings of teraploid crossing diploid,the offsprings of triploid crossing diploid,and the self-progenies of triploid.These polyploidy germplasms laid the important foundation for breeding polyploid apple cultivars.
引文
1.巴拉诺夫著,鲍文奎译.1959.植物多倍体.北京:科学出版社.
2.蔡旭.1988.植物遗传育种学.北京:科学出版社,596.
3.常金华,罗耀武.1997.高粱同源四倍体与二倍体胚胎发育的比较Ⅱ—胚及胚乳发育的比较.河北农业大学学报,20(4):6-10.
4.晁无疾,段丛.1985.去壁低渗法在葡萄染色体观察上的应用.葡萄栽培与酿酒,(3):1-3.
5.陈俊,李登科,李太保.1997.葡萄多倍体化学诱发育种方法.中外葡萄与葡萄酒,(2):26-27.
6.陈俊,唐晓萍,李登科.2001.早熟大粒优质葡萄新品种一早黑宝.园艺学报,28(3):277.
7.陈立松,万蜀渊.1996.一个新的桠柑突变体一'桂林良丰'无核柑的细胞学鉴定.武汉植物学研究,14(1):1-5.
8.陈士炎,叶大鹏.1989.茶树倍性与保卫细胞叶绿体数目的关系.茶叶科学,9(2):127-132.
9.陈学森,郭延奎.1991.几种果树多倍体与二倍体梢端细胞组织学的研究.山东农业大学学报,22(4):35-36.
10.陈绪中,罗正荣.2005.鄂柿1号离体叶片秋水仙素诱导和再生植株倍性鉴定.果树学报,22(5):554-556.
11.成明昊.1987.苹果属新种—小金海棠.农业科技通讯,9:19.
12.程心旻,高山林,卞云云.2003.白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较.植物资源与环境学报,12(1):16-20.
13.代红艳.2007.山楂种质资源的分子鉴定及创新研究.沈阳:沈阳农业大学.
14.戴洪义.1990.葡萄的染色体倍性与气孔性状的关系及其判别分析.葡萄栽培与酿酒,2:5-9
15.邓秀新,Fred G Gmitter J,Grosser JW.1995.柑桔同源及异源四倍体花粉育性研究.园艺学报,22(1):16-20.
16.邓秀新,Grosser JM,Gmitter FG.1995.柑桔种间体配融合及培养研究.遗传学报,22(4):316-321.
17.方从兵.2002.我国果树孢粉学研究进展(综述).安徽农业大学学报,29(2):154-157.
18.冯德江,刘翔,朱祯.2003.转录后基因沉默—植物抵御外来病毒入侵的一种机制.遗传学报,30(6):589-596.
19.谷晓峰,罗正荣.2003.秋水仙素处理'罗田甜柿'获得12倍体再生植株.园艺学报,30(3):325-327.
20.谷晓峰,罗正荣.2003.柿品种禅寺丸的花粉染色体加倍研究.中国农业科学,36(4):426-428.
21.郭海滨,刘向东,卢永根.2006.同源四倍体水稻成熟胚囊的结构及异常现象.中国水稻科学,20(3):283-289.
22.郭启高,宋明,梁国鲁.2000.植物多倍体诱导育种研究进展.生物学通报,35(2):8-10.
23.韩礼星,赵改荣,李玉红,张海漩,樊秀彩.1998.猕猴桃多倍体诱导研究.果树科学,15:273-276.
24.贺超兴,徐炳卢.1991.苹果属花粉形态特征及其分类学和进化意义.植物分类学报.29(5):445-451.
25.洪柳,邓秀新.2005.应用MSAP技术对脐橙品种进行DNA甲基化分析.中国农业科学,38(11):2306-2307.
26.黄礼森,李树玲,丛佩华.1990.梨多倍体与二倍体性状比较.中国果树,3:30-31.
27.黄群策,秦广雍,梁秋霞.2004.同源四倍体水稻与狼尾草杂交结实的胚胎学证据.杂交水稻,19(1):56-59.
28.黄群策,向茂成,汤国雄.2002.同源四倍体水稻与假稻杂交结籽的胚胎学机理研究.作物学报,28(2):286-288.
29.黄荣峰,郭培路,黄大防.2001.植物DNA甲基化.生物技术通报,5:1-3.
30.胡楷.2002.外因遗传学及其重要意义.遗传,6:23-29.
31.景士西.2000.园艺植物育种总论.北京:中国农业出版社.
32.李世诚,金佩芳,蒋爱丽.1998.与四倍体杂交的无核葡萄胚珠培养获得三倍体植株.上海农业学报,14(4):13-17.
33.李树玲,曹玉芬,黄礼森.1998.大鸭梨与不同倍性梨品种杂交后代染色体数目鉴定.天津农学院学报,5(4):1-7.
34.李树贤.2003.植物同源多倍体育种的几个问题.西北植物学报,23(10):1829-1841.
35.李旭刚,朱祯,冯德江,常团结,刘翔.2001.DNA甲基化对转基因表达的影响.科学通报,46(4):322-326.
36.李育农.2001.苹果属植物种质资源研究.北京:中国农业出版.
37.李赞,石荫坪,束怀瑞.1998.利用苹果花粉粒形态进行倍性鉴定.园艺学报,25(2):133-138.
38.李赞,石荫坪,束怀瑞.1999.应用气孔性状对苹果与梨的倍性判别分析.果树科学,16(1):9-13.
39.李赟,束怀瑞,石荫坪.1999.苹果三倍体和二倍体细胞超微结构观察.西北农业学报,8(3):68-70.
40.李云,姜金仲.2005.饲料型四倍体刺槐引种现状.东北林业大学学报,32(Suppl.):137-139.
41.李云,朱之悌,田砚亭,张志毅,康向阳.2001.秋水仙碱处理白杨雌花芽培育三倍体植株的研究.林业科学,37(5):68-74.
42.梁国鲁,李育农,李晓林.1996.中国苹果属植物小孢子减数分裂染色体系统研究.西南农业大学学报,18(4):299-307.
43.粱国鲁,李晓林.1993.中国苹果属植物染色体研究.植物分类学报,31(3):236-251.
44.廖乾初,蓝芬兰编著.1990.扫描电镜原理及应用技术.治金工业出版社.
45.刘惠吉,王华,高丽红.1997.不同生态型同源四倍体和二倍体白菜品种的同功酶比较.南京农业大学学报,20(3):26-30.
46.刘静,丁兆堂,赵进红,孙海伟,孙仲序.2003.茶树多倍体诱变研究初报.山东农业大学学报(自 然科学版),34(4):475-478.
47.刘庆忠,刘鹏,赵红军,孙玉刚.2002.同源四倍体皇家嘎啦苹果的生物学及光合生理特性研究.中国农业科学,35(12):1573-1578.
48.刘庆忠,赵红军,刘鹏,Mong Rengong,Freddi Hammerschlag.2001.秋水仙素处理离体叶片获得皇家嘎拉四倍体植株.果树学报,18(1):7-10.
49.刘仁祥,黄莺.1998.烟草花粉植株染色体倍性的早期快速鉴定.贵州农业科学,(6):4-7.
50.刘文革,王鸣,阎志红.2004.西瓜二倍体及同源多倍体遗传差异的AFLP分析.果树学报,21(1):46-49.
51.栾丽,孔繁伦,何涛,刘刚,图胜斌.2004.用SSR标记检测同源四倍体与二倍体水稻的遗传差异.应用与环境生物学报,10(5):556-558.
52.罗言云,贾勇炯,曹有龙,刘继明.1999.银杏成熟胚的离体培养研究.四川大学学报(自然科学版),363:573-576.
53.马国斌,王鸣,郑学勤.1999.甜瓜组织培养再生植株中的四倍体变异.园艺学报,26(2):128-130.
54.潘学军,张文娥,唐冬梅,骆强伟.2007.无核葡萄离体快繁技术研究.中国南方果树,36(2),44-46
55.裴新澍.1963.多倍体诱导与育种.上海:上海科学技术出版社.
56.乔永刚和宋芸.2002.我国园艺植物多倍体育种研究进展.北方园艺,(6):7-8.
57.瞿素萍,熊丽,莫锡君,王继华,张颢,苏艳.2004.香石竹的多倍体诱导及其变异研究.西南农业大学学报(自然科学版),26(5):609-612.
58.山东省果树研究所资源室.1982.几种果树花粉电镜扫描观察.山东果树,(4):27.
59.沈德绪,王元裕,陈力耕.1998.柑橘遗传育种学.北京:科学出版社,161-166.
60.沈德绪著.1995.果树育种学(第二版).北京:中国农业出版社,107-117.
61.石荫坪,李雅志,王强生.1986.果树突变育种.上海:上海技术出版社.
62.石荫坪,王强生,贾元淑.1985.枣胚乳三倍体的细胞学.中国果树资源细胞学之六.武汉植物学报,3(4):389-396.
63.石荫坪,王强生,尹永胜.1983.大玫瑰香葡萄的细胞学和育种行为.山东农学院学报,8:1-6.
64.石荫坪,王强生.1983.果树的多倍体.园艺,(2):6-12.
65.束怀瑞.1999.苹果学.北京:中国农业出版社.
66.谭德冠,庄南生,黄华孙.2005.组织培养与秋水仙碱诱导相结合培育植物多倍体的应用(综述).亚热带植物科学,34(1):77-80.
67.唐仙英,罗正荣.2000.部分中国柿品种及其胚培养后代的染色体倍性研究.园艺学报,27:235-239.
68.汪卫星,郭启高,粱国鲁.2003.热激处理对枇杷2n花粉发生率的影响.果树学报,20(4):284-286.
69.王长泉,张文胜,李雅志,崔得才.1999.苹果叶片离体培养中秋水仙素加倍效应的研究.果树科学,16(2):104-109.
70.王建波,张文驹.2000.异源多倍体植物核rDNA序列的同步进化.遗传,22:54-56.
71.王兰,刘向东,卢永根,冯九焕,徐雪宾,徐是雄.2004.同源四倍体水稻胚乳发育:糊粉层细胞壁纤维素物质发育、胚乳淀粉积累及胼胝质“套”的形成.中国水稻科学,18(6):28-32.
72.王兰,刘向东,卢永根.2004.同源四倍体水稻胚乳发育:极核融合和胚乳细胞化.中国水稻科学,18(4):281-289.
73.王娜,刘孟军,代丽,秦子禹.2005.秋水仙素离体诱导冬枣和酸枣四倍体.园艺学报,32(6):1008-1012.
74.王强生.1987.利用花粉鉴定果树多倍性种质的研究.植物学会交流,176:11-37.
75.王同坤,张京政,齐永顺,庞海珍.2004.我国果树多倍体育种研究进展.果树学报,21(6):592-597.
76.王卓伟,余茂德,鲁成.2002.桑树二倍体及人工诱导的同源四倍体遗传差异的AFLP分析.植物学通报,19(2):194-200.
77.吴丽杰,李甲斌.2002.苹果二倍体和三倍体几个生理指标的比较.河北果树,1:4-5.
78.吴筱丹,王华,楼键.2005.白菜DNA甲基化水平的反相离子对高效液相色谱法研究.理化检验,41:5-10.
79.吴雅琴,赵艳华,李春敏.2006.昌红苹果离体叶片不定芽的诱导及植株再生.云南农业大学学报,21(1):32-35.
80.武振华,牛炳韬,王新宇.2005.药用植物染色体加倍的研究进展.西北植物学报,25(12):2569-2574.
81.谢小群,高山林.2002.黄芩过氧化物酶同工酶电泳和抗坏血酸过氧化物酶活性分析.植物资源与环境学报,11(1):5-8.
82.Mirautadze NG(薛玉柱译).1989.γ射线照射茶籽后生成的多倍体类型.茶叶文摘,(4):No.890528.
83.徐德聪,吕芳德,潘晓杰.2003.叶绿素荧光分析技术在果树研究中的应用.经济林研究,21(3):88-91.
84.徐喜楼,徐惠瑛,盛炳成,程宗明,戴文浩.1985.苹果属花粉扫描电镜观察.南京农业大学学报,2(2):124-129.
85.闫忠业,伊凯,李作轩,刘志,王冬梅,杨锋.2006.富士芽变系品种花粉形态初探.园艺学报33(6):1299-1302.
86.杨传友,史金玉,杜欣阁,董会,苗良.1998.苹果叶片气孔的研究.山东农业大学学报(自然科学版),29(1):8-14.
87.杨继.2001.植物多倍体基因组的形成与进化.植物分类学报,39(4):357-371.
88.杨晓红,李育农.1995.北美绿苹果组植物的花粉形态和系统学研究.西南农业大学学报, 17(1):18-23.
89.杨晓红,李育农.1995.苹果属植物苹果组及山荆子组花粉形态及其演化研究.西南农业大学学报,17(4):279-285.
90.杨晓红,李育农.1995.苹果属植物中花楸苹果组和多胜海棠组的花粉形态和系统学研究.西南农业大学学报,17(4):348-354.
91.杨晓红,林培均.1992.新疆野苹果Malus sieversii(Ldb.)Roem花粉形态及其起源演化研究.西南农业大学学报,14(1):45-50.
92.杨晓红.1986.苹果属植物花粉观察研究.西南农业大学学报,2:121-129.
93.杨晓红.1995.世界苹果属植物花粉扫描电镜观察分析.西南农业大学学报.17(4):325-354.
94.张红宇,彭海,李云,徐培洲,汪旭东,吴先军2006.水稻基因组DNA胞嘧啶甲基化在单倍体和对应二倍体间的差异.科学通报,51(13):1529-1535.
95.张华华,冯九焕,卢永根,杨秉耀,刘向东.2003.利用激光扫描共聚焦显微镜观察同源四倍体水稻胚囊的形成发育.电子显微学报,22(5):380-384.
96.张华华,刘向东,卢永根,杨秉耀,刘向东.2006.同源四倍体水稻受精与胚胎形成过程的观察.激光生物学报,15(1):9-14.
97.张全美,张明芳.2003.园艺植物多倍体诱导研究进展.细胞生物学杂志,(4):223-228.
98.张文樾.1982.白香蕉葡萄芽变系‘吉香'的细胞学研究.北方园艺,4:1-5.
99.张新忠,刘国俭,章德明.1998.热激处理对桃、李2n花粉产生的影响.园艺学报,25(3):292-293.
100.赵辉,戴祖云.2006.西瓜多倍体育种研究进展.长江蔬菜,9:30-32.
101.赵卫国,苗雪霞,黄勇平,潘一乐.2005.桑树二倍体及其同源四倍体遗传差异的ISSR分析.蚕业科学,31(4):393-397.
102.周广芳,王成强.1997.果树多倍体育种途径与成就.落叶果树,(增刊):50-52.
103.周元昌,S.Kennedy.2002.抱子甘蓝花培苗倍性的快速鉴定.福建农林大学学报(自然科学版),31(1):55-58.
104.周祉祯.1979.干燥处理诱发蚕豆根尖细胞多倍体化.遗传学报,6(1):50.
105.Adams KL,Wendel JF.2003.Polyploidy and genome evolution in plants.Curr Opin Plant Biol,8:135-141.
106.Adelberg JW,Rhodes BB,Skorupska H T.1993.Generating tetraploid melons in tissue culture.Acta Hort,336:373-380.
107.Al-Janabi SM,McClelland M,Petersen C,Sobr BWS.1994.Phylogenetic analysis of organellar DNA sequences in the Andropo goneas:Saccharum.Theor.Appl.Genet,88:933-944.
108.Burge GK,Morgan ER,Seelye JF.2002.Opportunities for synthetic plant chimeral breeding:past and fruture.Plant Cell,tissue and Organ Clture,70:13-21.
109.Cao XF,Jacobsen SE.2002.Locus specific control of asymmetric and CpNpGmethylation by the DRM and CMT3 methyltransferase genes.Proc Natl Acad Sci USA, 99:16491—16498.
110.Cervera M T,Ruiz-Garcia L,Martinez-Zapater J M.2002.Analysis of DNA methylation in Arabidopsis thaliana based on methylation-sensitive AFLP markers.Molecular and General Genomics,268(4): 543-552.
111.Choi MY.1997.Improvement of chloroplast observation technique in guard cells for ploidy detection of microspore derived plants in broccoli.Journal of the Korean Society for Horticultural Science, 38 (6): 666-669.
112.Comai L, Tyagi AP, Winter K, Holmes Davis R, Reynolds SH, Stevens Y, Byers B.2000.Phenotypic instability and rapid gene silencing in newly formed Arabidopsis allotetraploids.Plant Cell, 12: 1551-1568.
113.Comai L.2000.Genetic and epigenetic interactions in allopolyploid plants.Plant Mol Biol, 43: 387—399.
114.Compton M, BaraettN, Gray D.1999.Use of fluorescein diacetate( FAD) to determine ploidy of in vitro watermelon shoots .Plant Cell, Tissue and Organ Culture, 58(3): 199—203 .
115.Cubas P, Vincent C, Coen E.1999.An epigenetic mutation responsible for natural variation in floral symmetry.Nature, 401:157—161.
116.Demming Adams B, Adam SW.1992.Photoprotection and other response of plants to high light stress.Annual Review of Plant Physiology and Plant Molecular Biology, 43 (1): 59 9—622.
117.Duren M, Morpurgo R, Dolezel J, Afza R.1996.Induction and verification of autotetraploids in diploid banana (Musa acuminata) by in vitro techniques.Euphytica, 88(1): 25—34.
118.Dover G A.1994.Concerted evolution: molecular drive and natural selection.Curt Biol, 4:1165.
119.Einset J.1952.Spontaneous polyploidy in cultivated apples.Amer Soc.HortScience, 59: 291—302.
120.Elosta A.2004.Coordination of epigenetic events.Cellular and Molecular Life Science, 17:2135—2136.
121.Feldman.M, Liu, Segal G, Abbo S, Levy AA, Vega JM.1997.Rapid elimination of low copy DNA sequences in polyploid wheat.Genetics, 147:1381—1387.
122.Finnegan FJ, Kovac KA.2000.Plant DNA methyltransferases.PlantMolecular Biology, 43:189—201.123.Fogle HW.1977.Identification of tree fruit species by pollen ultra structure.J Amer Soc Hort Sci,102 (5): 548-551.
124.Fogle HW.1977.Identification clones with in four tree fruit species by pollen exine patterns.J Amer Soc Hort Sci, 102 (5): 552-560.
125.Guillaume Blanc, Kenneth H Wolfe.2004.Functional divergence of duplicated genes formed by polyploidy during arabidopsis evolution.The Plant Cell, 16:1679—1691.
126.Ho I, Wan Y, Widholm JM.1990.The use of stomata chloroplast number for rapid determination of ploidy level in maize.Plant Breeding, 105(3): 203—210.
127.Kadota M, Niimi Y.2002.In vitro induction of tetraploid plants from a diploid Japanese pear cultivar (Pyrus pyrifolia N.cv.Hosui).Plant Cell Reports, 21(3): 282-286.
128.Kaneyoshi J, Kano T, Kuwata Y, Hirao A, Nakatani S, Kobayashi S.1997.Breeding of triploid citrus cultivars.I.production of triploids from Satsuma Mandarin (Citrus unshiu Marc.) X Tetraploid Ponkan Mandarin (Citrus reticulata Blanco) crosses.Journal of the Japanese Society for Horticultural Science, 66(1): 9～14.
129.King BJ, Lee LS, Scott PT.1996.Identification of triploid Citrus by isozyme analysis.Euphytica, 90(2): 223-231.
130.Koul KK, Raina SN.1996.Male and female meiosis in diploid and colchitetraploid Phlox dummondii Hook.(Polemoniaceae).Bot .J.Linnean.Soc, 122: 243—251.
131.Laubscher FX, Hurter N.1960.The chromosome numbers of seedling progeny from triploid apples.S African J Sci, 3: 31—39.
132.Liu B, Wendel JF.2003.Epigenetic phenomena and the evolution of plant allopolyploids.Mol Phylogenet Evol, 29(3):365～379.
133.Long S P.1994.Ann Rev Plant Physiol Plant Mol Biol, 45: 633.
134.Luo ZW, Hackett CA, Bradshaw JE, McNicol JW, Milbourne D.2001.Construction of a genetic linkage map in tetraploid species using molecular markers.Genetics(Bethesda), 157(3): 1369.
135.Madlung A, Masuelli RW, Watson B, Reynolds SH, Davison J, Comai L.2002.Remodeling of DNA methylation and phenotypic and transcriptional changes in synthetic arabidopsis allotetraploids.Plant Physiol, 129 (2): 733-746.
136.Marcucci MC, Sansavini S, Ciampolini F, Cresti M.1984.Distinguishing apple clone and cultivars by surface morphology and pollen physiology.J Amer Soc Hort Sci, 109 (1): 10—19.
137.Martienssen RA, Colot V.2001.DNA methylation and epigenetic inheritance in plants and filamentous fungi.Science, 293: 1070—1074.
138.Masterson J.1994.Stomatal size in fossil plants evidence for polyploidy in majority of angiosperms.Science, 264: 421—424.
139.Nilsson EH.1938.Production of forest trees with increased chromosome number and increased timber yield.Svensk Papp Tidn, (2): 5.
140.Ohgawara T, Kobayashi S, Ohgawara E, Uchimiya H, Ishii S.1985.Somatic hybrid plants obtained by protoplast fusion between Citrus sinensis and Poncirus trifoliate.TAG Theoretical and Applied Genetics,71(1): 1432—2242
141.Ohmi C, Wakana A, Shiraishi S.1993.Study of the parentage of grape cultivars by genetic interpretation of GPI-2 and PGM-2 isozymes.Euphytica, 65:195—221.
142.Peggy OA, Robert LJ.1994.Nuclear DNA content and ploidy levels in the genus ipomoe.J.Amer.Soc.Hort Sci., 119(1): 110-115.
143.Portis E.Acquadro A,Comino C,Lanteri S.2004.Analysis of DNA methylation during germination of peper ( Capsicum annuurn L.) seeds using methylation-sensitive amplification polymorphism (MSAP) .Plant Science, 166:169-178.
144.Quist G, Chow W, Anderson JM.1992.Photoinhibtion ofPhotosynthesis Rep resents aMechanism for the Longterm Regulation of Photosystemll.Planta, 186 (6): 450—460.
145.Rieseberg LH.2001.Polyploid evolution: keeping the peace at genomic reunions.Curr Biol, 13:925-928.
146.Rieseberg LH, Raymond O, RosenthalDM.2003.Major ecological transitions in annual sunflowers facilitated by hybridization.Science, 301:1211— 1216.
147.Rosenthal DM, Schwarzbach AE, Donovan LA, Raymond O, Rieseberg LH.2002.Phenotypic differentiation between three ancient hybrid taxa and their parental species.International Journal ofPlant Science (Chicago), 163(3): 387-401.
148.Salina EA, Numerova OM, Ozkan H, Feldman M.2004.Alterations in subtelomeric tandem repeats during early stages of allopolyplody in wheat.Genome, 47(5): 860-867.
149.Schranz ME, Osborn TC.2000.Novel flowering time variation in the resynthesized polyploid Bra ssica napus.J Hered, 91(3): 242—246.
150.Shaked H, Kashkush K, Ozkan H, Feldman M, Levy A.2001.A.Sequence elimination and cytosine methylation are rapid and reproducible responses of the genome to wide hybridization and allopolyploidy in wheat.The Plant Cell, 13(8): 1749-1759.
151.Shao JZ, Chen CL, Deng XX.2003.In vitro induction of tetraploid in pomegranate (Punica granatum).Plant Cell, Tissue and Organ Culture, 75(3): 241-246.
152.Shi Y, Wang Q, Zhou G, Wang J.1992.Genome engineering of apple in vitro.Acta Hort., 317: 13—21.
153.Stebbins GL.1971.Chromosomal evolution in higher piant.Edward Arnold, London.
154.Sugawara K, Oowada A.1995.Identification of Citrus chimeras by RAPD markers.Hortscience, 30: 1276-1278.
155.Sugawara K, Wakizuka T, Owada A.2002.Histogenic identification by RAPD analysis of leaves and fruit of newly synthesized chimeric citrus.J Amer Soc HortSci, 127(1): 104—107.
156.Tatum TC, Stepanovic S, Biradar DP, Rayburn AL, Korban SS.2005.Variation in nuclear DNA content in Malus species and cultivated apples.Genome, 48: 924—930.
157.Vandenhout H, Ortiz R, Vaylstteke D.1995.Effect of ploidy on stomatal and other quantitative traits in plantain and banana hybrids.Euphytica, 83:117—192.
158.Wendel JF.2000.Genome evolution in polyploids.Plant Mol Biol, 42(1): 225—249.
159.Xiong LZ, Xu CG, Maroof MA, Zhang QF.1999.Patterns of cytosine methylation in an elite rice hybrid and its parental lines,detected by a methylation-sensitive amplification polymorphism technique.Molecular and General Genetics, 261: 439～446.
160.Yang XM, Cao ZY, An LZ, Wang YM, Fang XW.2006.In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.).Euphytica, 152(2): 217-224.
161.Zhou JM, Hirata Y, Nou IS, Shiotani H, Ito T.2002.Interactions between different genotypic tissues in citrus graft chimeras.Euphytica, 126: 355 ～ 364.
2.蔡旭.1988.植物遗传育种学.北京:科学出版社,596.
3.常金华,罗耀武.1997.高粱同源四倍体与二倍体胚胎发育的比较Ⅱ—胚及胚乳发育的比较.河北农业大学学报,20(4):6-10.
4.晁无疾,段丛.1985.去壁低渗法在葡萄染色体观察上的应用.葡萄栽培与酿酒,(3):1-3.
5.陈俊,李登科,李太保.1997.葡萄多倍体化学诱发育种方法.中外葡萄与葡萄酒,(2):26-27.
6.陈俊,唐晓萍,李登科.2001.早熟大粒优质葡萄新品种一早黑宝.园艺学报,28(3):277.
7.陈立松,万蜀渊.1996.一个新的桠柑突变体一'桂林良丰'无核柑的细胞学鉴定.武汉植物学研究,14(1):1-5.
8.陈士炎,叶大鹏.1989.茶树倍性与保卫细胞叶绿体数目的关系.茶叶科学,9(2):127-132.
9.陈学森,郭延奎.1991.几种果树多倍体与二倍体梢端细胞组织学的研究.山东农业大学学报,22(4):35-36.
10.陈绪中,罗正荣.2005.鄂柿1号离体叶片秋水仙素诱导和再生植株倍性鉴定.果树学报,22(5):554-556.
11.成明昊.1987.苹果属新种—小金海棠.农业科技通讯,9:19.
12.程心旻,高山林,卞云云.2003.白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较.植物资源与环境学报,12(1):16-20.
13.代红艳.2007.山楂种质资源的分子鉴定及创新研究.沈阳:沈阳农业大学.
14.戴洪义.1990.葡萄的染色体倍性与气孔性状的关系及其判别分析.葡萄栽培与酿酒,2:5-9
15.邓秀新,Fred G Gmitter J,Grosser JW.1995.柑桔同源及异源四倍体花粉育性研究.园艺学报,22(1):16-20.
16.邓秀新,Grosser JM,Gmitter FG.1995.柑桔种间体配融合及培养研究.遗传学报,22(4):316-321.
17.方从兵.2002.我国果树孢粉学研究进展(综述).安徽农业大学学报,29(2):154-157.
18.冯德江,刘翔,朱祯.2003.转录后基因沉默—植物抵御外来病毒入侵的一种机制.遗传学报,30(6):589-596.
19.谷晓峰,罗正荣.2003.秋水仙素处理'罗田甜柿'获得12倍体再生植株.园艺学报,30(3):325-327.
20.谷晓峰,罗正荣.2003.柿品种禅寺丸的花粉染色体加倍研究.中国农业科学,36(4):426-428.
21.郭海滨,刘向东,卢永根.2006.同源四倍体水稻成熟胚囊的结构及异常现象.中国水稻科学,20(3):283-289.
22.郭启高,宋明,梁国鲁.2000.植物多倍体诱导育种研究进展.生物学通报,35(2):8-10.
23.韩礼星,赵改荣,李玉红,张海漩,樊秀彩.1998.猕猴桃多倍体诱导研究.果树科学,15:273-276.
24.贺超兴,徐炳卢.1991.苹果属花粉形态特征及其分类学和进化意义.植物分类学报.29(5):445-451.
25.洪柳,邓秀新.2005.应用MSAP技术对脐橙品种进行DNA甲基化分析.中国农业科学,38(11):2306-2307.
26.黄礼森,李树玲,丛佩华.1990.梨多倍体与二倍体性状比较.中国果树,3:30-31.
27.黄群策,秦广雍,梁秋霞.2004.同源四倍体水稻与狼尾草杂交结实的胚胎学证据.杂交水稻,19(1):56-59.
28.黄群策,向茂成,汤国雄.2002.同源四倍体水稻与假稻杂交结籽的胚胎学机理研究.作物学报,28(2):286-288.
29.黄荣峰,郭培路,黄大防.2001.植物DNA甲基化.生物技术通报,5:1-3.
30.胡楷.2002.外因遗传学及其重要意义.遗传,6:23-29.
31.景士西.2000.园艺植物育种总论.北京:中国农业出版社.
32.李世诚,金佩芳,蒋爱丽.1998.与四倍体杂交的无核葡萄胚珠培养获得三倍体植株.上海农业学报,14(4):13-17.
33.李树玲,曹玉芬,黄礼森.1998.大鸭梨与不同倍性梨品种杂交后代染色体数目鉴定.天津农学院学报,5(4):1-7.
34.李树贤.2003.植物同源多倍体育种的几个问题.西北植物学报,23(10):1829-1841.
35.李旭刚,朱祯,冯德江,常团结,刘翔.2001.DNA甲基化对转基因表达的影响.科学通报,46(4):322-326.
36.李育农.2001.苹果属植物种质资源研究.北京:中国农业出版.
37.李赞,石荫坪,束怀瑞.1998.利用苹果花粉粒形态进行倍性鉴定.园艺学报,25(2):133-138.
38.李赞,石荫坪,束怀瑞.1999.应用气孔性状对苹果与梨的倍性判别分析.果树科学,16(1):9-13.
39.李赟,束怀瑞,石荫坪.1999.苹果三倍体和二倍体细胞超微结构观察.西北农业学报,8(3):68-70.
40.李云,姜金仲.2005.饲料型四倍体刺槐引种现状.东北林业大学学报,32(Suppl.):137-139.
41.李云,朱之悌,田砚亭,张志毅,康向阳.2001.秋水仙碱处理白杨雌花芽培育三倍体植株的研究.林业科学,37(5):68-74.
42.梁国鲁,李育农,李晓林.1996.中国苹果属植物小孢子减数分裂染色体系统研究.西南农业大学学报,18(4):299-307.
43.粱国鲁,李晓林.1993.中国苹果属植物染色体研究.植物分类学报,31(3):236-251.
44.廖乾初,蓝芬兰编著.1990.扫描电镜原理及应用技术.治金工业出版社.
45.刘惠吉,王华,高丽红.1997.不同生态型同源四倍体和二倍体白菜品种的同功酶比较.南京农业大学学报,20(3):26-30.
46.刘静,丁兆堂,赵进红,孙海伟,孙仲序.2003.茶树多倍体诱变研究初报.山东农业大学学报(自 然科学版),34(4):475-478.
47.刘庆忠,刘鹏,赵红军,孙玉刚.2002.同源四倍体皇家嘎啦苹果的生物学及光合生理特性研究.中国农业科学,35(12):1573-1578.
48.刘庆忠,赵红军,刘鹏,Mong Rengong,Freddi Hammerschlag.2001.秋水仙素处理离体叶片获得皇家嘎拉四倍体植株.果树学报,18(1):7-10.
49.刘仁祥,黄莺.1998.烟草花粉植株染色体倍性的早期快速鉴定.贵州农业科学,(6):4-7.
50.刘文革,王鸣,阎志红.2004.西瓜二倍体及同源多倍体遗传差异的AFLP分析.果树学报,21(1):46-49.
51.栾丽,孔繁伦,何涛,刘刚,图胜斌.2004.用SSR标记检测同源四倍体与二倍体水稻的遗传差异.应用与环境生物学报,10(5):556-558.
52.罗言云,贾勇炯,曹有龙,刘继明.1999.银杏成熟胚的离体培养研究.四川大学学报(自然科学版),363:573-576.
53.马国斌,王鸣,郑学勤.1999.甜瓜组织培养再生植株中的四倍体变异.园艺学报,26(2):128-130.
54.潘学军,张文娥,唐冬梅,骆强伟.2007.无核葡萄离体快繁技术研究.中国南方果树,36(2),44-46
55.裴新澍.1963.多倍体诱导与育种.上海:上海科学技术出版社.
56.乔永刚和宋芸.2002.我国园艺植物多倍体育种研究进展.北方园艺,(6):7-8.
57.瞿素萍,熊丽,莫锡君,王继华,张颢,苏艳.2004.香石竹的多倍体诱导及其变异研究.西南农业大学学报(自然科学版),26(5):609-612.
58.山东省果树研究所资源室.1982.几种果树花粉电镜扫描观察.山东果树,(4):27.
59.沈德绪,王元裕,陈力耕.1998.柑橘遗传育种学.北京:科学出版社,161-166.
60.沈德绪著.1995.果树育种学(第二版).北京:中国农业出版社,107-117.
61.石荫坪,李雅志,王强生.1986.果树突变育种.上海:上海技术出版社.
62.石荫坪,王强生,贾元淑.1985.枣胚乳三倍体的细胞学.中国果树资源细胞学之六.武汉植物学报,3(4):389-396.
63.石荫坪,王强生,尹永胜.1983.大玫瑰香葡萄的细胞学和育种行为.山东农学院学报,8:1-6.
64.石荫坪,王强生.1983.果树的多倍体.园艺,(2):6-12.
65.束怀瑞.1999.苹果学.北京:中国农业出版社.
66.谭德冠,庄南生,黄华孙.2005.组织培养与秋水仙碱诱导相结合培育植物多倍体的应用(综述).亚热带植物科学,34(1):77-80.
67.唐仙英,罗正荣.2000.部分中国柿品种及其胚培养后代的染色体倍性研究.园艺学报,27:235-239.
68.汪卫星,郭启高,粱国鲁.2003.热激处理对枇杷2n花粉发生率的影响.果树学报,20(4):284-286.
69.王长泉,张文胜,李雅志,崔得才.1999.苹果叶片离体培养中秋水仙素加倍效应的研究.果树科学,16(2):104-109.
70.王建波,张文驹.2000.异源多倍体植物核rDNA序列的同步进化.遗传,22:54-56.
71.王兰,刘向东,卢永根,冯九焕,徐雪宾,徐是雄.2004.同源四倍体水稻胚乳发育:糊粉层细胞壁纤维素物质发育、胚乳淀粉积累及胼胝质“套”的形成.中国水稻科学,18(6):28-32.
72.王兰,刘向东,卢永根.2004.同源四倍体水稻胚乳发育:极核融合和胚乳细胞化.中国水稻科学,18(4):281-289.
73.王娜,刘孟军,代丽,秦子禹.2005.秋水仙素离体诱导冬枣和酸枣四倍体.园艺学报,32(6):1008-1012.
74.王强生.1987.利用花粉鉴定果树多倍性种质的研究.植物学会交流,176:11-37.
75.王同坤,张京政,齐永顺,庞海珍.2004.我国果树多倍体育种研究进展.果树学报,21(6):592-597.
76.王卓伟,余茂德,鲁成.2002.桑树二倍体及人工诱导的同源四倍体遗传差异的AFLP分析.植物学通报,19(2):194-200.
77.吴丽杰,李甲斌.2002.苹果二倍体和三倍体几个生理指标的比较.河北果树,1:4-5.
78.吴筱丹,王华,楼键.2005.白菜DNA甲基化水平的反相离子对高效液相色谱法研究.理化检验,41:5-10.
79.吴雅琴,赵艳华,李春敏.2006.昌红苹果离体叶片不定芽的诱导及植株再生.云南农业大学学报,21(1):32-35.
80.武振华,牛炳韬,王新宇.2005.药用植物染色体加倍的研究进展.西北植物学报,25(12):2569-2574.
81.谢小群,高山林.2002.黄芩过氧化物酶同工酶电泳和抗坏血酸过氧化物酶活性分析.植物资源与环境学报,11(1):5-8.
82.Mirautadze NG(薛玉柱译).1989.γ射线照射茶籽后生成的多倍体类型.茶叶文摘,(4):No.890528.
83.徐德聪,吕芳德,潘晓杰.2003.叶绿素荧光分析技术在果树研究中的应用.经济林研究,21(3):88-91.
84.徐喜楼,徐惠瑛,盛炳成,程宗明,戴文浩.1985.苹果属花粉扫描电镜观察.南京农业大学学报,2(2):124-129.
85.闫忠业,伊凯,李作轩,刘志,王冬梅,杨锋.2006.富士芽变系品种花粉形态初探.园艺学报33(6):1299-1302.
86.杨传友,史金玉,杜欣阁,董会,苗良.1998.苹果叶片气孔的研究.山东农业大学学报(自然科学版),29(1):8-14.
87.杨继.2001.植物多倍体基因组的形成与进化.植物分类学报,39(4):357-371.
88.杨晓红,李育农.1995.北美绿苹果组植物的花粉形态和系统学研究.西南农业大学学报, 17(1):18-23.
89.杨晓红,李育农.1995.苹果属植物苹果组及山荆子组花粉形态及其演化研究.西南农业大学学报,17(4):279-285.
90.杨晓红,李育农.1995.苹果属植物中花楸苹果组和多胜海棠组的花粉形态和系统学研究.西南农业大学学报,17(4):348-354.
91.杨晓红,林培均.1992.新疆野苹果Malus sieversii(Ldb.)Roem花粉形态及其起源演化研究.西南农业大学学报,14(1):45-50.
92.杨晓红.1986.苹果属植物花粉观察研究.西南农业大学学报,2:121-129.
93.杨晓红.1995.世界苹果属植物花粉扫描电镜观察分析.西南农业大学学报.17(4):325-354.
94.张红宇,彭海,李云,徐培洲,汪旭东,吴先军2006.水稻基因组DNA胞嘧啶甲基化在单倍体和对应二倍体间的差异.科学通报,51(13):1529-1535.
95.张华华,冯九焕,卢永根,杨秉耀,刘向东.2003.利用激光扫描共聚焦显微镜观察同源四倍体水稻胚囊的形成发育.电子显微学报,22(5):380-384.
96.张华华,刘向东,卢永根,杨秉耀,刘向东.2006.同源四倍体水稻受精与胚胎形成过程的观察.激光生物学报,15(1):9-14.
97.张全美,张明芳.2003.园艺植物多倍体诱导研究进展.细胞生物学杂志,(4):223-228.
98.张文樾.1982.白香蕉葡萄芽变系‘吉香'的细胞学研究.北方园艺,4:1-5.
99.张新忠,刘国俭,章德明.1998.热激处理对桃、李2n花粉产生的影响.园艺学报,25(3):292-293.
100.赵辉,戴祖云.2006.西瓜多倍体育种研究进展.长江蔬菜,9:30-32.
101.赵卫国,苗雪霞,黄勇平,潘一乐.2005.桑树二倍体及其同源四倍体遗传差异的ISSR分析.蚕业科学,31(4):393-397.
102.周广芳,王成强.1997.果树多倍体育种途径与成就.落叶果树,(增刊):50-52.
103.周元昌,S.Kennedy.2002.抱子甘蓝花培苗倍性的快速鉴定.福建农林大学学报(自然科学版),31(1):55-58.
104.周祉祯.1979.干燥处理诱发蚕豆根尖细胞多倍体化.遗传学报,6(1):50.
105.Adams KL,Wendel JF.2003.Polyploidy and genome evolution in plants.Curr Opin Plant Biol,8:135-141.
106.Adelberg JW,Rhodes BB,Skorupska H T.1993.Generating tetraploid melons in tissue culture.Acta Hort,336:373-380.
107.Al-Janabi SM,McClelland M,Petersen C,Sobr BWS.1994.Phylogenetic analysis of organellar DNA sequences in the Andropo goneas:Saccharum.Theor.Appl.Genet,88:933-944.
108.Burge GK,Morgan ER,Seelye JF.2002.Opportunities for synthetic plant chimeral breeding:past and fruture.Plant Cell,tissue and Organ Clture,70:13-21.
109.Cao XF,Jacobsen SE.2002.Locus specific control of asymmetric and CpNpGmethylation by the DRM and CMT3 methyltransferase genes.Proc Natl Acad Sci USA, 99:16491—16498.
110.Cervera M T,Ruiz-Garcia L,Martinez-Zapater J M.2002.Analysis of DNA methylation in Arabidopsis thaliana based on methylation-sensitive AFLP markers.Molecular and General Genomics,268(4): 543-552.
111.Choi MY.1997.Improvement of chloroplast observation technique in guard cells for ploidy detection of microspore derived plants in broccoli.Journal of the Korean Society for Horticultural Science, 38 (6): 666-669.
112.Comai L, Tyagi AP, Winter K, Holmes Davis R, Reynolds SH, Stevens Y, Byers B.2000.Phenotypic instability and rapid gene silencing in newly formed Arabidopsis allotetraploids.Plant Cell, 12: 1551-1568.
113.Comai L.2000.Genetic and epigenetic interactions in allopolyploid plants.Plant Mol Biol, 43: 387—399.
114.Compton M, BaraettN, Gray D.1999.Use of fluorescein diacetate( FAD) to determine ploidy of in vitro watermelon shoots .Plant Cell, Tissue and Organ Culture, 58(3): 199—203 .
115.Cubas P, Vincent C, Coen E.1999.An epigenetic mutation responsible for natural variation in floral symmetry.Nature, 401:157—161.
116.Demming Adams B, Adam SW.1992.Photoprotection and other response of plants to high light stress.Annual Review of Plant Physiology and Plant Molecular Biology, 43 (1): 59 9—622.
117.Duren M, Morpurgo R, Dolezel J, Afza R.1996.Induction and verification of autotetraploids in diploid banana (Musa acuminata) by in vitro techniques.Euphytica, 88(1): 25—34.
118.Dover G A.1994.Concerted evolution: molecular drive and natural selection.Curt Biol, 4:1165.
119.Einset J.1952.Spontaneous polyploidy in cultivated apples.Amer Soc.HortScience, 59: 291—302.
120.Elosta A.2004.Coordination of epigenetic events.Cellular and Molecular Life Science, 17:2135—2136.
121.Feldman.M, Liu, Segal G, Abbo S, Levy AA, Vega JM.1997.Rapid elimination of low copy DNA sequences in polyploid wheat.Genetics, 147:1381—1387.
122.Finnegan FJ, Kovac KA.2000.Plant DNA methyltransferases.PlantMolecular Biology, 43:189—201.123.Fogle HW.1977.Identification of tree fruit species by pollen ultra structure.J Amer Soc Hort Sci,102 (5): 548-551.
124.Fogle HW.1977.Identification clones with in four tree fruit species by pollen exine patterns.J Amer Soc Hort Sci, 102 (5): 552-560.
125.Guillaume Blanc, Kenneth H Wolfe.2004.Functional divergence of duplicated genes formed by polyploidy during arabidopsis evolution.The Plant Cell, 16:1679—1691.
126.Ho I, Wan Y, Widholm JM.1990.The use of stomata chloroplast number for rapid determination of ploidy level in maize.Plant Breeding, 105(3): 203—210.
127.Kadota M, Niimi Y.2002.In vitro induction of tetraploid plants from a diploid Japanese pear cultivar (Pyrus pyrifolia N.cv.Hosui).Plant Cell Reports, 21(3): 282-286.
128.Kaneyoshi J, Kano T, Kuwata Y, Hirao A, Nakatani S, Kobayashi S.1997.Breeding of triploid citrus cultivars.I.production of triploids from Satsuma Mandarin (Citrus unshiu Marc.) X Tetraploid Ponkan Mandarin (Citrus reticulata Blanco) crosses.Journal of the Japanese Society for Horticultural Science, 66(1): 9～14.
129.King BJ, Lee LS, Scott PT.1996.Identification of triploid Citrus by isozyme analysis.Euphytica, 90(2): 223-231.
130.Koul KK, Raina SN.1996.Male and female meiosis in diploid and colchitetraploid Phlox dummondii Hook.(Polemoniaceae).Bot .J.Linnean.Soc, 122: 243—251.
131.Laubscher FX, Hurter N.1960.The chromosome numbers of seedling progeny from triploid apples.S African J Sci, 3: 31—39.
132.Liu B, Wendel JF.2003.Epigenetic phenomena and the evolution of plant allopolyploids.Mol Phylogenet Evol, 29(3):365～379.
133.Long S P.1994.Ann Rev Plant Physiol Plant Mol Biol, 45: 633.
134.Luo ZW, Hackett CA, Bradshaw JE, McNicol JW, Milbourne D.2001.Construction of a genetic linkage map in tetraploid species using molecular markers.Genetics(Bethesda), 157(3): 1369.
135.Madlung A, Masuelli RW, Watson B, Reynolds SH, Davison J, Comai L.2002.Remodeling of DNA methylation and phenotypic and transcriptional changes in synthetic arabidopsis allotetraploids.Plant Physiol, 129 (2): 733-746.
136.Marcucci MC, Sansavini S, Ciampolini F, Cresti M.1984.Distinguishing apple clone and cultivars by surface morphology and pollen physiology.J Amer Soc Hort Sci, 109 (1): 10—19.
137.Martienssen RA, Colot V.2001.DNA methylation and epigenetic inheritance in plants and filamentous fungi.Science, 293: 1070—1074.
138.Masterson J.1994.Stomatal size in fossil plants evidence for polyploidy in majority of angiosperms.Science, 264: 421—424.
139.Nilsson EH.1938.Production of forest trees with increased chromosome number and increased timber yield.Svensk Papp Tidn, (2): 5.
140.Ohgawara T, Kobayashi S, Ohgawara E, Uchimiya H, Ishii S.1985.Somatic hybrid plants obtained by protoplast fusion between Citrus sinensis and Poncirus trifoliate.TAG Theoretical and Applied Genetics,71(1): 1432—2242
141.Ohmi C, Wakana A, Shiraishi S.1993.Study of the parentage of grape cultivars by genetic interpretation of GPI-2 and PGM-2 isozymes.Euphytica, 65:195—221.
142.Peggy OA, Robert LJ.1994.Nuclear DNA content and ploidy levels in the genus ipomoe.J.Amer.Soc.Hort Sci., 119(1): 110-115.
143.Portis E.Acquadro A,Comino C,Lanteri S.2004.Analysis of DNA methylation during germination of peper ( Capsicum annuurn L.) seeds using methylation-sensitive amplification polymorphism (MSAP) .Plant Science, 166:169-178.
144.Quist G, Chow W, Anderson JM.1992.Photoinhibtion ofPhotosynthesis Rep resents aMechanism for the Longterm Regulation of Photosystemll.Planta, 186 (6): 450—460.
145.Rieseberg LH.2001.Polyploid evolution: keeping the peace at genomic reunions.Curr Biol, 13:925-928.
146.Rieseberg LH, Raymond O, RosenthalDM.2003.Major ecological transitions in annual sunflowers facilitated by hybridization.Science, 301:1211— 1216.
147.Rosenthal DM, Schwarzbach AE, Donovan LA, Raymond O, Rieseberg LH.2002.Phenotypic differentiation between three ancient hybrid taxa and their parental species.International Journal ofPlant Science (Chicago), 163(3): 387-401.
148.Salina EA, Numerova OM, Ozkan H, Feldman M.2004.Alterations in subtelomeric tandem repeats during early stages of allopolyplody in wheat.Genome, 47(5): 860-867.
149.Schranz ME, Osborn TC.2000.Novel flowering time variation in the resynthesized polyploid Bra ssica napus.J Hered, 91(3): 242—246.
150.Shaked H, Kashkush K, Ozkan H, Feldman M, Levy A.2001.A.Sequence elimination and cytosine methylation are rapid and reproducible responses of the genome to wide hybridization and allopolyploidy in wheat.The Plant Cell, 13(8): 1749-1759.
151.Shao JZ, Chen CL, Deng XX.2003.In vitro induction of tetraploid in pomegranate (Punica granatum).Plant Cell, Tissue and Organ Culture, 75(3): 241-246.
152.Shi Y, Wang Q, Zhou G, Wang J.1992.Genome engineering of apple in vitro.Acta Hort., 317: 13—21.
153.Stebbins GL.1971.Chromosomal evolution in higher piant.Edward Arnold, London.
154.Sugawara K, Oowada A.1995.Identification of Citrus chimeras by RAPD markers.Hortscience, 30: 1276-1278.
155.Sugawara K, Wakizuka T, Owada A.2002.Histogenic identification by RAPD analysis of leaves and fruit of newly synthesized chimeric citrus.J Amer Soc HortSci, 127(1): 104—107.
156.Tatum TC, Stepanovic S, Biradar DP, Rayburn AL, Korban SS.2005.Variation in nuclear DNA content in Malus species and cultivated apples.Genome, 48: 924—930.
157.Vandenhout H, Ortiz R, Vaylstteke D.1995.Effect of ploidy on stomatal and other quantitative traits in plantain and banana hybrids.Euphytica, 83:117—192.
158.Wendel JF.2000.Genome evolution in polyploids.Plant Mol Biol, 42(1): 225—249.
159.Xiong LZ, Xu CG, Maroof MA, Zhang QF.1999.Patterns of cytosine methylation in an elite rice hybrid and its parental lines,detected by a methylation-sensitive amplification polymorphism technique.Molecular and General Genetics, 261: 439～446.
160.Yang XM, Cao ZY, An LZ, Wang YM, Fang XW.2006.In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.).Euphytica, 152(2): 217-224.
161.Zhou JM, Hirata Y, Nou IS, Shiotani H, Ito T.2002.Interactions between different genotypic tissues in citrus graft chimeras.Euphytica, 126: 355 ～ 364.