本文選題：萱草 + 種間雜交��； 參考：《沈陽農業(yè)大學》2017年博士論文

【摘要】：萱草(Hemerocallis spp.)花型花色豐富多樣,抗性強、適應性好,被廣泛用作園林綠化。萱草育種經過一百多年的研究,品種已達8萬以上,但由于長期品種間的雜交導致萱草品種遺傳基礎日趨狹窄,而種間雜交是萱草品種遺傳改良、新品種選育的一個重要途徑。本研究利用我國東北特有野生萱草資源與品種進行雜交,獲得種間雜交后代,并對萱草屬植物種間雜交親和性及雜交障礙克服進行了系統(tǒng)研究;花為萱草的主要觀賞部位,本研究還對雜交后代花性狀的遺傳變異進行分析,同時對不同花色的萱草屬植物進行花瓣色素成分初步定性分析,以期為萱草屬植物種間雜交的花性狀遺傳改良及遺傳變異研究提供科學依據。主要結果如下:利用液體培養(yǎng)基法測定萱草品種'玫瑰回旋'的花粉生活力,最佳培養(yǎng)條件是25℃C、3h,最佳培養(yǎng)基是100 g·L-1蔗糖+60 mg·L-1硼酸+40mg·L-1氯化鈣�；ǚ凵盍ψ罡叩氖嵌扼w野生種北黃花菜(H lilioasphodelus),達到64.54%,最低的為三倍體野生種萱草(H.fulva),為28.28%。柱頭可授性測定結果表明,除'金娃娃'在開花前一天上午9:00具有最強柱頭可授性以外,其他5種萱草均在開花當天上午9:00具有最強的柱頭可授性,且持續(xù)時間最強的是'回復'。不同倍性萱草屬植物種間雜交親和性研究顯示:二倍體萱草種間雜交果實的平均膨大率為48.02%,結實率為31.57%,單果種子數平均為9.39個,種子出芽率為80.07%,后代幼苗成活率為60.94%,白化苗率為3.70%。重瓣萱草的后代白化苗率最高,平均為10.47%。二倍體萱草與四倍體萱草的種間雜交及三倍體萱草作父本與二倍體萱草做母本的種間雜交均不結實,三倍體萱草做母本與二倍體萱草的種間雜交結實率平均為19.58%,后代出芽率35.00%,幼苗成活率38.71%,白化苗率為6.35%。萱草屬植物種間雜交障礙主要包括:受精前障礙和受精后障礙。受精前的萱草花粉母細胞減數分裂觀察,發(fā)現有多種染色體發(fā)生異常行為現象,包括不等二價體、染色體橋和片段、滯后染色體、微核、三分體和多分體等異常染色體,這些都會導致不正常的花粉出現,從而影響育性。種間雜交花粉管熒光觀察發(fā)現存在許多異常生長現象,如花粉在柱頭上不萌發(fā),萌發(fā)卻不繼續(xù)生長,生長卻不進入柱頭,花粉管生長進入花柱卻纏繞和回折不能到達子房,到達到子房卻不進入胚珠等;授粉后的胚胎學觀察發(fā)現,胚胎發(fā)育異�，F象主要表現在:胚囊未進行雙受精,胚與胚乳發(fā)育滯后,胚乳正常發(fā)育而胚停留在球形胚階段并退化,胚正常而無胚乳等。為了克服雜交障礙,主要方法包括:應用超低溫-80℃燥貯藏花粉克服花期不遇;應用不同授粉方式解決受精前雜交障礙,重復授粉可以平均增加10%的后代雜交結實率,0.1%NAA涂抹對果實膨大率和雜交結實率均有極顯著的促進;采用離體培養(yǎng)幼胚的方式解決受精后雜交障礙,胚齡30d以后的離體培養(yǎng)萌發(fā)率高達90%以上,授粉后40d的幼胚離體培養(yǎng)萌發(fā)率可達100%,雜交種胚培養(yǎng)最適的培養(yǎng)基是MS+NAA0.1 mg·L-1 +6-BA2 mg·L-1。對萱草屬種間雜交親本及后代的花性狀進行遺傳變異分析,獲得了 20%花直徑顯著高于高親的雜交后代,最大可達12.20 cm。在'回復'做母本的雜交后代中有10.00%的植株花數/序大于17.7個,最多的可達20.33個。以'重瓣魏河'為母本與三種單瓣花萱草品種進行雜交,后代中平均花瓣數量均大于6個,但是重瓣花后代所占的比例不同,花瓣數量最多的是重瓣魏河×粉邊玫紅的后代DF-11,平均花瓣數量為12個。對花期的研究發(fā)現,二次抽花序特性可以顯著延長萱草的單株花期和群體花期,具有二次抽花序特性的親本作母本與不具有二次抽花序特性的親本雜交,后代中均含有二次抽花序的植株。通過目測、比色卡和色差儀的測量,將萱草屬種間雜交后代的花色劃分為3個色系:黃色系(黃色和橙色)、粉色系和紅色系�；ㄉz傳規(guī)律表明,紅色系的'回復'作親本,無論是與黃色、橙色還是粉色系親本雜交,后代紅色花所占比例均較高,說明紅色花的遺傳力較強。另外,種間雜交后代的花色分布較廣,說明萱草的花色遺傳可能是由多基因控制的。對4種顏色的萱草花瓣結構與色素分布進行了研究,黃色和橙色萱草色素在花瓣上表皮、下表皮和中間的柵欄組織均有分布,粉色和紅色萱草色素主要分布在花瓣上表皮與下表皮的細胞中。深紅色萱草花瓣上表皮有明顯的圓錐形突起,加大了對光線的吸收。對4種顏色的萱草屬植物花瓣的色素成分進行了初步定性分析,認為萱草花瓣中主要含有4種色素成分,包括花青素苷、類胡蘿卜素、黃酮類化合物和黃酮醇類化合物,具有綠色喉的萱草花瓣中含有少量葉綠素。
[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.
【學位授予單位】：沈陽農業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S682.19

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