萱草属植物种间杂交亲和性及花性状遗传研究

发布时间：2018-07-02 13:02

本文选题：萱草 + 种间杂交　；参考：《沈阳农业大学》2017年博士论文

【摘要】：萱草(Hemerocallis spp.)花型花色丰富多样,抗性强、适应性好,被广泛用作园林绿化。萱草育种经过一百多年的研究,品种已达8万以上,但由于长期品种间的杂交导致萱草品种遗传基础日趋狭窄,而种间杂交是萱草品种遗传改良、新品种选育的一个重要途径。本研究利用我国东北特有野生萱草资源与品种进行杂交,获得种间杂交后代,并对萱草属植物种间杂交亲和性及杂交障碍克服进行了系统研究;花为萱草的主要观赏部位,本研究还对杂交后代花性状的遗传变异进行分析,同时对不同花色的萱草属植物进行花瓣色素成分初步定性分析,以期为萱草属植物种间杂交的花性状遗传改良及遗传变异研究提供科学依据。主要结果如下:利用液体培养基法测定萱草品种'玫瑰回旋'的花粉生活力,最佳培养条件是25℃C、3h,最佳培养基是100 g·L-1蔗糖+60 mg·L-1硼酸+40mg·L-1氯化钙。花粉生活力最高的是二倍体野生种北黄花菜(H lilioasphodelus),达到64.54%,最低的为三倍体野生种萱草(H.fulva),为28.28%。柱头可授性测定结果表明,除'金娃娃'在开花前一天上午9:00具有最强柱头可授性以外,其他5种萱草均在开花当天上午9:00具有最强的柱头可授性,且持续时间最强的是'回复'。不同倍性萱草属植物种间杂交亲和性研究显示:二倍体萱草种间杂交果实的平均膨大率为48.02%,结实率为31.57%,单果种子数平均为9.39个,种子出芽率为80.07%,后代幼苗成活率为60.94%,白化苗率为3.70%。重瓣萱草的后代白化苗率最高,平均为10.47%。二倍体萱草与四倍体萱草的种间杂交及三倍体萱草作父本与二倍体萱草做母本的种间杂交均不结实,三倍体萱草做母本与二倍体萱草的种间杂交结实率平均为19.58%,后代出芽率35.00%,幼苗成活率38.71%,白化苗率为6.35%。萱草属植物种间杂交障碍主要包括:受精前障碍和受精后障碍。受精前的萱草花粉母细胞减数分裂观察,发现有多种染色体发生异常行为现象,包括不等二价体、染色体桥和片段、滞后染色体、微核、三分体和多分体等异常染色体,这些都会导致不正常的花粉出现,从而影响育性。种间杂交花粉管荧光观察发现存在许多异常生长现象,如花粉在柱头上不萌发,萌发却不继续生长,生长却不进入柱头,花粉管生长进入花柱却缠绕和回折不能到达子房,到达到子房却不进入胚珠等;授粉后的胚胎学观察发现,胚胎发育异常现象主要表现在:胚囊未进行双受精,胚与胚乳发育滞后,胚乳正常发育而胚停留在球形胚阶段并退化,胚正常而无胚乳等。为了克服杂交障碍,主要方法包括:应用超低温-80℃燥贮藏花粉克服花期不遇;应用不同授粉方式解决受精前杂交障碍,重复授粉可以平均增加10%的后代杂交结实率,0.1%NAA涂抹对果实膨大率和杂交结实率均有极显著的促进;采用离体培养幼胚的方式解决受精后杂交障碍,胚龄30d以后的离体培养萌发率高达90%以上,授粉后40d的幼胚离体培养萌发率可达100%,杂交种胚培养最适的培养基是MS+NAA0.1 mg·L-1 +6-BA2 mg·L-1。对萱草属种间杂交亲本及后代的花性状进行遗传变异分析,获得了 20%花直径显著高于高亲的杂交后代,最大可达12.20 cm。在'回复'做母本的杂交后代中有10.00%的植株花数/序大于17.7个,最多的可达20.33个。以'重瓣魏河'为母本与三种单瓣花萱草品种进行杂交,后代中平均花瓣数量均大于6个,但是重瓣花后代所占的比例不同,花瓣数量最多的是重瓣魏河×粉边玫红的后代DF-11,平均花瓣数量为12个。对花期的研究发现,二次抽花序特性可以显著延长萱草的单株花期和群体花期,具有二次抽花序特性的亲本作母本与不具有二次抽花序特性的亲本杂交,后代中均含有二次抽花序的植株。通过目测、比色卡和色差仪的测量,将萱草属种间杂交后代的花色划分为3个色系:黄色系(黄色和橙色)、粉色系和红色系。花色遗传规律表明,红色系的'回复'作亲本,无论是与黄色、橙色还是粉色系亲本杂交,后代红色花所占比例均较高,说明红色花的遗传力较强。另外,种间杂交后代的花色分布较广,说明萱草的花色遗传可能是由多基因控制的。对4种颜色的萱草花瓣结构与色素分布进行了研究,黄色和橙色萱草色素在花瓣上表皮、下表皮和中间的栅栏组织均有分布,粉色和红色萱草色素主要分布在花瓣上表皮与下表皮的细胞中。深红色萱草花瓣上表皮有明显的圆锥形突起,加大了对光线的吸收。对4种颜色的萱草属植物花瓣的色素成分进行了初步定性分析,认为萱草花瓣中主要含有4种色素成分,包括花青素苷、类胡萝卜素、黄酮类化合物和黄酮醇类化合物,具有绿色喉的萱草花瓣中含有少量叶绿素。
[Abstract]:Hemerocallis spp. (Hemerocallis Hemerocallis (Hemerocallis Hemerocallis) has a wide variety of flower color, strong resistance and good adaptability. It has been widely used as garden greening. After more than 100 years of study on Hemerocallis Hemerocallis, the variety has reached more than 80 thousand. However, the genetic foundation of Hemerocallis Hemerocallis is becoming narrower because of the hybridization between long varieties, and interspecific hybridization is the genetic improvement of Hemerocallis Hemerocallis and the selection of new varieties. An important way. In this study, the hybrid progeny of interspecific hybrids was obtained from the wild Hemerocallis Hemerocallis in the northeast of China, and the Interspecific Hybridization Compatibility and overcoming of hybrids were systematically studied. The flower is the main ornamental part of the Hemerocallis, and the genetic variation of the flower characters of the hybrids was also divided. In order to provide scientific basis for the genetic improvement and genetic variation of flower traits in the interspecific hybrids of the genus Hemerocallis, the main results are as follows: the main results are as follows: the determination of the pollen viability and the optimum conditions of the "Rose cyclotron" by the liquid culture base method. It is 25 C, 3h, the best medium is 100 g. L-1 sucrose +60 mg. L-1 boric acid +40mg. L-1 calcium chloride. The highest pollen viability is the diploid wild species of North Yellow Flower (H lilioasphodelus), reaching 64.54%, the lowest is the triploid wild species of Hemerocallis Hemerocallis (H.fulva). In addition to the strongest stigma receptivity at 9:00, the other 5 kinds of Hemerocallis had the strongest stigma receptivity at 9:00 a.m. on the day of flowering, and the strongest duration was' recovery '. The study on the Interspecific Hybridization Compatibility of different hybrids showed that the average expansion rate of the diploid hybrids was 48.02%, the seed setting rate was 31.57%, single fruit. The average seed number is 9.39, the seed germination rate is 80.07%, the survival rate of the offspring is 60.94%, the albino seedling rate is the highest in the offspring of 3.70%. heavy Hemerocallis Hemerocallis, and the interspecific hybridization between the 10.47%. diploid Hemerocallis Hemerocallis and the tetraploid Hemerocallis Hemerocallis and the triploid of Hemerocallis Hemerocallis and the diploid Hemerocallis Hemerocallis are not strong and triploid. The interspecific seed setting rate of Hemerocallis Hemerocallis and diploid Hemerocallis Hemerocallis is 19.58%, the germination rate of descendants is 35%, the survival rate of the seedlings is 38.71%. The interspecific hybridity obstacle of Hemerocallis Hemerocallis 6.35%. is mainly included: pre fertilization barrier and post fertilization barrier. Abnormal behavior, including unequal two valence body, chromosome bridge and fragment, lagging chromosome, micronucleus, three body and multibody and other abnormal chromosomes, all of which lead to abnormal pollen appearance and affect fertility. The pollen tube fluorescence observation of interspecific hybridization found that there are many abnormal growth phenomena, such as pollen germination on stigma, and germination of pollen. But it does not continue to grow, but the growth does not enter the stigma, the pollen tube grows into the style but does not reach the ovary, but does not enter the ovary, but does not enter the ovary. After pollination, the embryological observation found that the abnormal embryo development is mainly manifested in the unfertilized embryo sac, the development of the embryo and the endosperm, the normal development of the endosperm and the stay of the embryo. In order to overcome hybridism, the main methods include: using ultra-low temperature -80 centigrade storage pollen to overcome the flowering period, and using different pollination methods to solve the cross barrier before fertilization. Repeated pollination can increase the rate of 10% later hybrid seed setting, and 0.1%NAA smear on fruit enlargement rate and impurity. The rate of crossbreeding was greatly promoted. The germination rate of the isolated embryo after fertilization was up to 90% after the embryo age of 30d, and the germination rate of the young embryos in vitro was up to 100% after pollination, and the optimum medium for the hybrid embryo culture was MS +NAA0.1 mg. L-1 +6-BA2 mg. L-1. to the interspecies of Hemerocallis. Genetic variation analysis was carried out on the flower traits of the parents and offspring, and the 20% flowers with a diameter of 20% flowers were significantly higher than those of the high parent. The maximum number of 12.20 cm. in the hybrid offspring of the 'recovery' parent was more than 17.7, up to 20.33. The "double petal Wei River" was the mother and the three single flower Hemerocallis. The average petal number in the offspring was more than 6, but the proportion of the petals was different, the most of the petals was the offspring of the double petal Wei River and the pink edge rose DF-11, and the average petal number was 12. The study of florescence found that the two inflorescence characteristics could significantly prolong the single plant and population flowering of Hemerocallis Hemerocallis, and have two pumping. The parent of the inflorescence was hybridized with the parent with no two inflorescence characteristics. The progeny of the progeny contained two plants of the inflorescence. The color of the hybrids of the hybrids of the genus Hemerocallis was divided into 3 colors by visual measurement, compared with the color card and chromatic aberration meter. The red line 'recovery' was used as parent, whether it was hybrid with yellow, orange or pink, the proportion of red flowers in the offspring was higher, indicating that the heritability of red flowers was stronger. In addition, the color distribution of interspecific hybrids was widely distributed, indicating that the color inheritance of Hemerocallis Hemerocallis may be controlled by multiple genes. The structure and color of the 4 color Hemerocallis flower petals The yellow and orange Hemerocallis Hemerocallis pigment is distributed in the upper epidermis, the lower epidermis and the middle palisade tissue. The pink and red Hemerocallis pigment is mainly distributed in the epidermis and the lower epidermis of the petals. The upper epidermis of the deep red Hemerocallis Hemerocallis petal has obvious conical protuberance, which increases the absorption of the light. 4 colors are used. The pigment composition of the flower petals of Hemerocallis is a preliminary qualitative analysis. It is considered that the main pigment in the flower petals of Hemerocallis Hemerocallis contains 4 kinds of pigment, including anthocyanins, carotenoids, flavonoids and flavonols, and the green larynx contains a small amount of lynx in the petals.
【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S682.19

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