【摘要】：射干(Iris domestica)與野鳶尾(I.dichotoma)同為鳶尾科(Iridaceze)鳶尾屬(Iris)植物,本試驗(yàn)通過(guò)射干與三種花色的野鳶尾進(jìn)行雜交鑒定射干的無(wú)融合生殖。射干分別與紫花野鳶尾、白花野鳶尾和黃花野鳶尾進(jìn)行正反交,獲得F1群體。從F1群體的每個(gè)組合中隨機(jī)選取20個(gè)株系進(jìn)行植物性性狀的調(diào)查。從射干做母本的三個(gè)組合中每個(gè)組合隨機(jī)選取4個(gè)株系進(jìn)行自交實(shí)驗(yàn),每個(gè)組合獲得80個(gè)株系,從80個(gè)株系中隨機(jī)選取20個(gè)株系進(jìn)行植物學(xué)性狀調(diào)查,分析射干做母本的三個(gè)組合的F2群體是否出現(xiàn)性狀分離。使用SNP(single nucleotide polymorphism,單核昔酸多態(tài)性)分子標(biāo)記法對(duì)射干與三種花色野鳶尾正反交后代的13個(gè)株系進(jìn)行了雜種真實(shí)性鑒定。使用花粉管熒光觀察和石蠟切片的方法觀察射干與野鳶尾雜交及射干自交過(guò)程中的花粉管行為和雙受精過(guò)程。結(jié)果表明,分別以三種花色野鳶尾做母本的雜交后代的植物學(xué)性狀表現(xiàn)為雙親的中間型,雜交后代的花色、花型、果子、種子等植物學(xué)性狀表現(xiàn)出和親本的差異性。以射干為母本與三種花色野鳶尾進(jìn)行雜交獲得的后代在植物學(xué)性狀上表現(xiàn)出偏母遺傳,雜交后代的花色、花型、果實(shí)以及種子均未表現(xiàn)出與母本的差異。在對(duì)射干做母本的雜交后代自交F2的植物學(xué)性狀調(diào)查后發(fā)現(xiàn),三個(gè)組合的F2群體并未出現(xiàn)性狀分離,三個(gè)組合的F2在花色、開(kāi)花時(shí)間、閉花時(shí)間、果實(shí)形態(tài)、種子形態(tài)性狀表現(xiàn)上均與其親本或射干差異不顯著。但在株高、花冠幅、內(nèi)外花被寬等一些數(shù)量性狀上,F2與其親本或射干差異顯著,不同組合的F2群體之間株高、花冠幅、內(nèi)外花被長(zhǎng)、內(nèi)外花被寬差異均不顯著。SNP分子標(biāo)記方法首次在鳶尾屬植物中成功應(yīng)用。紫花野鳶尾×射干的兩個(gè)株系、白花野鳶尾×射干的兩個(gè)株系和黃花野鳶尾×射干的三個(gè)株系均為真雜種,其雜交后代中既有來(lái)源于母本野鳶尾的SNP位點(diǎn),也有來(lái)源于父本射干的SNP位點(diǎn)。而射干×紫花野鳶尾、射干×白花野鳶尾、射干×黃花野鳶尾三個(gè)組合的六個(gè)株系均為假雜種,雜交后代只有來(lái)源于母本射干的SNP位點(diǎn),并沒(méi)有直接來(lái)源于父本野鳶尾的SNP位點(diǎn)。通過(guò)射干作為母本與三種花色野鳶尾雜交后代性狀一致且均有偏母特性、其雜交后代自交F2也未出現(xiàn)性狀分離的表現(xiàn)和SNP分子標(biāo)記法的鑒定結(jié)果為假雜種可以證明射干為無(wú)融合生殖,這是第一次在鳶尾屬植物中發(fā)現(xiàn)無(wú)融合生殖現(xiàn)象。對(duì)射干與野鳶尾雜交后的花粉管行為的觀察發(fā)現(xiàn),野鳶尾的花粉能夠在射干的柱頭上萌發(fā)伸長(zhǎng)到射干的胚珠內(nèi),但是出現(xiàn)了花粉管纏繞、萌發(fā)伸長(zhǎng)時(shí)間比射干自交要慢的現(xiàn)象。使用石蠟切片方法對(duì)射干自交和射干與野鳶尾雜交的雙受精過(guò)程進(jìn)行了觀察,射干自交在授粉后3d左右完成雙受精過(guò)程,并未觀察到射干與野鳶尾雜交完成雙受精的過(guò)程。
[Abstract]:Ejaculated (Iris domestica) and I.dichotoma are (Iris) plants of (Iridaceze) Iris family. In this experiment, the ejaculated apomixis was identified by hybridization with three species of wild iris. F _ 1 population was obtained by direct and negative crossing with wild iris of purple flower, wild iris of white flower and iris of yellow flower respectively. Twenty lines were randomly selected from each combination of F1 population to investigate the plant characters. Four lines were randomly selected from each of the three combinations to carry out self-crossing experiments. 80 lines were obtained from each combination, and 20 lines were randomly selected from 80 lines to investigate the botanical characters. The trait segregation of F 2 population with three combinations of ejaculation as female parent was analyzed. SNP (single nucleotide polymorphism, mononuclear acid polymorphism (SNP (single nucleotide polymorphism,) molecular marker method was used to identify the authenticity of 13 hybrids of Radix perforatum and three species of iris. The pollen tube behavior and the process of double fertilization were observed by fluorescence observation of pollen tube and paraffin section. The results showed that the botanical characters of hybrid progenies with three kinds of flower color wild iris as female parent were the middle type of parents, and the difference of botanical characters such as flower color, flower type, fruit, seed and so on of hybrid progeny showed difference with their parents. The results showed that the progeny of shoot dry as female parent and three kinds of flower color wild iris showed partial maternal inheritance in botanical characters, and the flower color, flower shape, fruit and seed of hybrid progeny showed no difference with female parent. After investigating the botanical characters of F _ 2 of female progeny, it was found that there was no segregation of characters in F2 population of three combinations, and the F _ 2 of three combinations were in flower color, flowering time, closing flower time and fruit morphology. There was no significant difference between seed morphological characters and their parents or ejaculation. But in some quantitative characters, such as plant height, Corolla width, width of inner and outer perianth, F2 had significant difference with their parents or shoot stem. The plant height, Corolla width, length of inner and outer perianth were different among F2 populations of different combinations. There was no significant difference in the width of perianth between internal and external flowers. SNP molecular marker method was successfully used in Iris for the first time. Two lines of Iris sinensis 脳 shoot stem, two lines of Iris albicans 脳 shoot dry and three strains of Iris huanghuaensis 脳 Radix perforated were true hybrids, and there were SNP loci derived from female iris in their hybrid progenies. There are also SNP loci derived from male ejaculation. The six lines of the three combinations were all pseudohybrids, and only the SNP loci derived from the female parent were found in the hybrid progenies, and the three combinations were all pseudohybrids, and the three combinations were all pseudohybrids, and the six lines of the three combinations were all pseudohybrids, and the hybrid progenies were only derived from the SNP loci of the female parent. There was no direct source of SNP loci from the male wild iris. By means of shoot dry as a female parent and three kinds of flower color wild iris hybrid progeny, the characters of the progenies are consistent and both have the characteristic of partial motherhood. There was no segregation of traits in F _ 2 and the result of SNP molecular marker analysis showed that the pseudo-hybrid could prove that the shoot stem was apomixis. This was the first time that apomixis was found in Iris. The pollen germination and elongation of the pollen of Iris wilfordii were found to be able to germinate and extend to the ovules on the stigma of the shoot, but the pollen tube was entangled, and the time of germination and elongation was slower than that of self-pollination in the pollen tube behavior after the crossing between the shoot and the iris, the pollen of the wild iris could germinate and extend to the ovules on the stigma of the shoot. The process of double fertilization was observed by using paraffin section method. The process of double fertilization was completed about 3 days after pollination, but the process of double fertilization was not observed.
【學(xué)位授予單位】：沈陽(yáng)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S682.19

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