發(fā)布時(shí)間：2018-01-02 01:20

  本文關(guān)鍵詞：木荷繁殖生物學(xué)特性及種子園交配系統(tǒng)研究　出處：《中國(guó)林業(yè)科學(xué)研究院》2017年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 木荷 種子園 繁殖生物學(xué) 自交不親和 交配系統(tǒng) 傳粉規(guī)律

【摘要】：木荷(Schima superba)為我國(guó)東部濕潤(rùn)亞熱帶常綠闊葉林的主要建群樹種,其材性優(yōu)良,抗逆性強(qiáng),是我國(guó)南方各省區(qū)中主要的珍貴優(yōu)質(zhì)闊葉用材和高效生物防火樹種。自2001年開展木荷良種選育以來,已在浙江、福建、江西、廣東和重慶等營(yíng)建了超過100ha的木荷無性系種子園。但對(duì)木荷的繁殖生物學(xué)特性和種子園的遺傳管理缺乏系統(tǒng)研究,難以為種子園的經(jīng)營(yíng)管理及良種生產(chǎn)提供理論和實(shí)踐指導(dǎo)。鑒于此,本文以進(jìn)入正常開花結(jié)實(shí)期的木荷種子園為研究對(duì)象,對(duì)木荷開花綜合特征、傳粉機(jī)制、繁育系統(tǒng)等進(jìn)行了深入研究,揭示木荷的繁殖生物學(xué)特性,為其種子生產(chǎn)、資源利用和雜交育種提供重要的實(shí)踐指導(dǎo)和科學(xué)依據(jù)。同時(shí),研究和揭示了種子園無性系花期同步性、遺傳交配系統(tǒng)狀況及傳粉規(guī)律,為木荷種子園營(yíng)建和高效的遺傳管理提供了重要的科學(xué)理論依據(jù)。主要研究結(jié)果如下:1.通過定點(diǎn)觀察,對(duì)木荷花部綜合特征及繁育系統(tǒng)的研究結(jié)果表明,木荷5~7月份開花,單花期4~5天�；ò咨�,常多朵排成總狀花序,平均每個(gè)花序著生7朵花。開花當(dāng)天柱頭即具有可授性,且在整個(gè)單花期內(nèi)均保持較高的可授性�；ǚ凼Щ钶^快,花粉活性在花朵剛開放時(shí)最高(83.78%),至花朵凋謝時(shí)花粉活性僅為5.45%�；ㄩ_后柱頭快速伸長(zhǎng),直至略高于花藥,形成柱頭和花藥在空間上的分離。木荷的雜交指數(shù)(OCI)值為4,花粉胚珠比(P/O)值約為6686.67,結(jié)合控制授粉試驗(yàn)顯示,木荷繁育系統(tǒng)以異交為主,不存在無融合生殖,傳粉過程需要傳粉者。2.木荷為異花授粉植物,其種子生產(chǎn)必須依靠昆蟲傳粉。通過觀察認(rèn)為,中華蜜蜂(Apis cerana)、白星花金龜(Protaetia brevitarsis)和棉花弧麗金龜(Popillia mutans)是木荷的主要傳粉昆蟲。中華蜜蜂和棉花弧麗金龜?shù)脑L花高峰均發(fā)生在10:00~11:00,白星花金龜則無明顯的訪花高峰。3種傳粉昆蟲體表均攜帶花粉,白星花金龜?shù)臄y粉量顯著高于棉花弧麗金龜(P0.001)和中華蜜蜂(P0.001),棉花弧麗金龜?shù)臄y粉量顯著高于中華蜜蜂(P0.001)。體表不同部位攜粉量不同,3種傳粉昆蟲主要的攜粉部位為胸腹部,同時(shí)也是接觸花藥和柱頭最為頻繁和力度最大的部位,因此它們的傳粉方式均為腹觸式傳粉。中華蜜蜂每次訪花的花粉移出數(shù)、柱頭花粉沉降數(shù)及傳粉者效率均低于白星花金龜和棉花弧麗金龜,但不顯著(P0.05),而中華蜜蜂在訪花頻率上占有明顯優(yōu)勢(shì),分別為白星花金龜?shù)?9倍和棉花弧麗金龜?shù)?.3倍。綜合各項(xiàng)特征表明,中華蜜蜂是木荷最有效的傳粉者。3.進(jìn)一步對(duì)木荷自交與異交的親和性進(jìn)行比較研究。結(jié)果顯示,自交與異交花粉均能在柱頭上正常萌發(fā),生長(zhǎng)至花柱基部。異交花粉管能成功進(jìn)入子房和胚囊,而自交花粉管不能進(jìn)入,表明木荷為晚期自交不親和(LSI)植物。自交和異交授粉均能引起子房SOD、POD和CAT活性的增加。自交和異交授粉48 h,氨基酸含量差異最大,其中自交授粉精氨酸(Arg)的含量顯著高于異交(P0.001),表明Arg含量的變化可能與木荷晚期自交不親和有關(guān)。授粉后子房生長(zhǎng)素(IAA)和玉米素(ZT)呈先升高后下降的趨勢(shì)。授粉48 h,異交子房IAA和ZT含量均顯著高于自交(P0.05),表明子房中高水平的IAA和ZT有利于授粉受精正常進(jìn)行。4.差異蛋白組學(xué)研究結(jié)果顯示,自交和異交授粉48 h后的子房存在82個(gè)差異蛋白點(diǎn),經(jīng)質(zhì)譜和數(shù)據(jù)庫檢索成功鑒定出58個(gè)蛋白點(diǎn),其中自交授粉中表達(dá)豐度升高的蛋白點(diǎn)有15個(gè),表達(dá)豐度降低的有43個(gè)。異交授粉后肌動(dòng)蛋白和蛋白酶體表達(dá)量顯著大于自交授粉,而過氧化物酶蛋白、蛋白質(zhì)二硫鍵異構(gòu)酶及植物凝集素蛋白的表達(dá)量顯著低于自交授粉。轉(zhuǎn)錄組測(cè)序結(jié)果顯示,自交與異交授粉48 h子房間篩選出509個(gè)差異表達(dá)基因,其中與信號(hào)轉(zhuǎn)導(dǎo)(Ca2+信號(hào)、蛋白激酶與蛋白磷酸酶等)、防御(細(xì)胞色素P450與MLP31等)、氨基酸代謝(草酰乙酸脫羧酶等)等相關(guān)的基因在自交子房中的表達(dá)量顯著高于異交,促進(jìn)生長(zhǎng)類激素代謝相關(guān)基因的表達(dá)量顯著低于異交。另外,自交子房中還發(fā)現(xiàn)2個(gè)特異表達(dá)的轉(zhuǎn)錄因子:MYB和bHLH,兩者共同參與調(diào)控木荷晚期自交不親和反應(yīng)。這些基因在轉(zhuǎn)錄組中表達(dá)結(jié)果與qRT-PCR分析結(jié)果一致。綜合分析表明,木荷晚期自交不親和反應(yīng)發(fā)生時(shí)伴隨著一系列的信號(hào)轉(zhuǎn)導(dǎo)與防衛(wèi)響應(yīng),是一個(gè)主動(dòng)抑制自交花粉管在子房中生長(zhǎng)的過程。5.連續(xù)2年對(duì)木荷種子園中19個(gè)無性系進(jìn)行花期、花量調(diào)查,分析了各無性系的開花物候特征和花期同步指數(shù)。相關(guān)分析結(jié)果表明,花量與花期長(zhǎng)度、座果數(shù)存在顯著正相關(guān)關(guān)系,花期長(zhǎng)度和座果數(shù)也存在顯著正相關(guān)關(guān)系。無性系組合間花期同步指數(shù)存在極顯著變異(P0.01),2015年和2016年的變化范圍分別為0.552~0.857和0.406~0.808,變異系數(shù)分別為12.016%~46.476%和15.375%~51.202%。2015年和2016年的平均花期同步指數(shù)分別為0.758和0.713,不存在顯著變異(P0.05)。年度間花期同步指數(shù)相關(guān)系數(shù)為0.229,表明年度間花期同步指數(shù)具有一定的相關(guān)性。種子園無性系年度間花期同步指數(shù)較為穩(wěn)定,可根據(jù)花期同步指數(shù)對(duì)無性系進(jìn)行篩選和優(yōu)化,同時(shí)輔以人工授粉等措施彌補(bǔ)花期同步性差異,以提高種子園種子產(chǎn)量和質(zhì)量。6.同時(shí),以木荷種子園投產(chǎn)初期自由授粉子代為研究對(duì)象,運(yùn)用13對(duì)SSR引物,對(duì)種子園內(nèi)44個(gè)親本及328個(gè)子代進(jìn)行分析。結(jié)果顯示,子代群體包含親本群體所有的等位基因。親本群體遺傳多樣性水平適中,子代群體遺傳多樣性較親本略有降低,觀測(cè)雜合度略高于親本,說明子代群體中實(shí)際觀察到的雜合單株的比例較親本有所增加,但差異不大。子代群體的F值為-0.143,存在雜合子過剩。多位點(diǎn)交配系統(tǒng)分析結(jié)果表明:種子園異交率較高,親本的近交現(xiàn)象不顯著,但有效花粉供體數(shù)目較少。單位點(diǎn)和多位點(diǎn)父本相關(guān)性的差值(0.012)大于0,表明只有小部分花粉供體是近親關(guān)系。11個(gè)家系間異交率差異不明顯,少數(shù)存在近交現(xiàn)象。11個(gè)家系的父本相關(guān)性和有效花粉供體數(shù)目變化較大,分別為0.210~0.762和1.3~4.8,說明各家系的父本相關(guān)性程度不一致,其中31號(hào)最高,最低的是48號(hào)。整體而言,木荷種子園異交率高,近交現(xiàn)象不明顯;無性系之間基因交流相對(duì)充分,遺傳多樣性豐富,子代能保持親本所具有的較高的遺傳多樣性。7.13對(duì)SSR引物在木荷種子園中檢測(cè)到親本和子代群體總的多態(tài)性信息含量(PIC)、觀測(cè)雜合度(Ho)和期望雜合度(He)分別為0.679、0.714和0.725。在95%的可信水平下總共鑒定得到203個(gè)全同胞家系,平均每個(gè)采種母樹與18個(gè)父本產(chǎn)生子代。在自由授粉狀態(tài)下,木荷無性系種子園自交率為1.5%,自交現(xiàn)象很弱,其交配方式以異交為主。父本繁殖貢獻(xiàn)率在0.49%~7.77%之間,平均為2.44%。木荷種子園的主要傳粉距離集中在0~60 m,檢測(cè)到的最大傳粉距離為120 m,其中0~60 m的傳粉距離內(nèi)產(chǎn)生的子代數(shù)量占已確定父本子代群體的80%以上。種子園存在一定的花粉污染,為7.01%。綜合分析認(rèn)為,木荷種子園花粉傳播較為均勻,種子園與花粉污染源隔離距離應(yīng)在60m以上,120 m以上最佳,以防止傳粉昆蟲的長(zhǎng)距離傳粉活動(dòng)造成花粉污染。
[Abstract]:Schimasuperba (Schima superba) the main constructive species in East China moist subtropical evergreen broad-leaved forests, its excellent material properties, strong resistance, is the main precious hardwood species and efficient biological fire provinces in the south of China. Since 2001 to carry out breeding in Schima superba has been in Zhejiang, Fujian, and Jiangxi. Guangdong and Chongqing construction of schimasuperba clonal seed orchard of more than 100HA. But the lack of systematic research on the genetic management of Schima superba reproductive characteristics and seed orchard, to provide theoretical and practical guidance for seed orchard management and seed production. In view of this, this paper to enter the normal flowering and fruiting stage of Schima superba seed orchard for research the object, flowering comprehensive characteristics, pollination mechanism of Schima superba, breeding system conducted in-depth research, reveal the reproductive characteristics of Schima superba, its seed production, utilization and breeding resources to provide heavy To practice guidance and scientific basis. At the same time, with the study of clonal seed orchard of flowering synchrony, genetic mating system and pollination, provides an important theoretical basis for the scientific management of Schima superba seed orchard establishment and genetic efficiency. The main results are as follows: 1. through the fixed-point observation research on wood, Lotus comprehensive department the characteristics and breeding system. The results showed that the flowering of Schima 5~7 month, single flower period of 4~5 days. Many white flowers, often arranged in racemes, inflorescences per 7 flowers. On the day of anthesis. With the receptivity, and maintain high receptivity in the single flower. Rapid inactivation of pollen the highest activity in pollen, just the flowers open (83.78%), to flower pollen activity is only 5.45%. flower stigma after rapid elongation, until slightly higher than anther, stigma and anther separation form in space. The hybrid index (wood bearing The OCI value is 4), pollen ovule ratio (P/O) value is about 6686.67 and the combination of controlled pollination experiments showed that Schima superba dominated by outcrossing breeding system, no apomixis, pollination of pollinator.2. Schima cross pollination, the seed production must be pollinated by insects. It has been observed that the bees (Apis, cerana) brevitarsis (Protaetia brevitarsis) and cotton beetle (Popillia mutans) arc is the main pollinators. The bees and Schima superba cotton arc beetle foraging peaks occurred at 10:00~11:00, white gold star turtle no flower peak.3 pollinators were obvious surface pollen carrying visit. Carrying amount of powder brevitarsis was significantly higher than that of cotton arc beetle (P0.001) and the Chinese honeybee (P0.001) powder, carrying amount of cotton was significantly higher than that of the arc beetle bees (P0.001). Different parts of the body with 3 kinds of different amount of powder, the main pollination insects To the site for carrying powder is also in contact with the chest and abdomen, anther and stigma is the most frequent and the largest part, so the pollination they were abdominal touch pollination. The number of each flower visiting Chinese bee pollen removal, pollen deposition and pollinator efficiency were lower than brevitarsis and cotton popillia beetles, but not significantly (P0.05), and the bee occupies obvious advantage in visitation frequency, were 8.3 times 29 times brevitarsis and cotton. The arc beetle showed that the characteristics of Chinese bee pollination is the most effective.3. schimasuperba further affinity for self and schimasuperba to make a comparative study. The results showed that the outcrossing and selfing pollen can germinate on the stigma, growth to base of style. Outcrossing pollen tube can successfully enter the ovary and embryo sac, and self pollen tube can not enter, show that the advanced self incompatibility schimasuperba (L SI). Plant selfing and outcrossing pollination can cause ovary SOD, increase POD and CAT activity. The self and cross pollinated 48 h, amino acid content of the biggest difference, the self pollination of arginine (Arg) were significantly higher than those of outcrossing (P0.001), indicating that the change of Arg content may be related to late wood bearing self incompatibility. Ovary after pollination, auxin (IAA) and Zeatin (ZT) increased firstly and then decreased. Pollination 48 h, outcrossing ovary IAA and ZT were significantly higher than that of self (P0.05), showed that in high levels of IAA and ZT is conducive to the normal pollination and fertilization by.4. protein study results show that the self and cross pollinated ovary after 48 h there are 82 differential protein spots by mass spectrometry and database retrieval successfully identified 58 proteins, including self pollination in up-regulated 15 proteins, expression decreased abundance was 43. Cross pollination after actin Protein and proteasome expression was significantly greater than that of self pollination, and the peroxidase protein expression of two protein disulfide isomerase and lectin protein was significantly lower than that of self pollination. Transcriptome sequencing results showed that the self pollination and cross pollination of 48 h sub room screened 509 differentially expressed genes, including Ca2+ (signal and signal transduction protein kinase and protein phosphatase, etc.), defense (cytochrome P450 and MLP31), amino acid metabolism (oxaloacetate decarboxylase etc.) and related gene expression in hybrids was significantly higher than that in the self promoting outcrossing, expression of growth hormone metabolism related genes was significantly lower than that of outcrossing. In addition, since the sub room also found 2 specific transcription factors: MYB and bHLH, both of them are involved in the regulation of late Schima superba self incompatibility. These gene expression results with qRT-PCR in comprehensive transcriptome. Analysis shows that the late schimasuperba self incompatibility occurs when accompanied by a series of signal transduction and defense response, is an active in inhibiting the growth of the ovary in the process of self pollen tube.5. for 2 consecutive years of 19 clones in seed orchard of Schima superba flowering, flower quantity survey, flowering phenology and synchronous flowering index analysis of the clones. The results of correlation analysis showed that the amount of flowers and flowering length, there is a significant positive correlation between fruit number, there are significant positive correlation between the length of flowering and fruit number. There is significant variation between clones combined flowering synchrony index (P0.01), the change range of 2015 and 2016 were 0.552~0.857 and 0.406~0.808, the coefficient of variation were 12.016%~46.476% and 15.375%~51.202%.2015 and 2016 average flowering synchrony index were 0.758 and 0.713, there is no significant variation (P0.05). The same year fl. Step index correlation coefficient was 0.229, showed that the annual flowering synchrony index has certain correlation. Clones in seed orchard of inter annual flowering synchronization index is relatively stable, according to the index of synchronous flowering clones were screened and optimized, supplemented by artificial pollination and other measures to compensate for the flowering synchrony differences, in order to improve the seed yield and quality of.6. Park seeds at the same time, to the early stage of production of Schima superba seed orchard open pollinated progeny as the research object, using 13 pairs of SSR primers on the seed orchard of 44 parents and 328 offspring were analyzed. The results showed that the offspring population contains parental populations all alleles. The genetic diversity level of moderate parental populations in progeny population, genetic diversity a parent was slightly reduced, the observed heterozygosity is slightly higher than that of the parents, shows the proportion of heterozygous individuals in progeny populations observed were increased, but the difference is small. The offspring group The F-measure is -0.143, there are heterozygote excess. Multilocus mating system analysis results showed that the seed orchard outcrossing rate is higher, the phenomenon of inbreeding were not significant, but the effective number of pollen donors. Less difference unit point and multilocus paternal correlation (0.012) is greater than 0, showed that only a small portion of the pollen donor.11 is a close relationship between families of outcrossing rates were not significantly different, there are few male parent pollen correlation and effective inbreeding.11 pedigrees of donor number changes greatly, respectively 0.210~0.762 and 1.3~4.8, the degree of correlation between male parent families is not a show, the 31 was highest, the lowest is 48 overall. In terms of Schima seed orchard high outcrossing rate, inbreeding phenomenon is not obvious; gene exchange between clones is relatively full, rich genetic diversity, genetic diversity of parent offspring can keep the higher.7.13 of SSR primers in Schima superba seed orchard To the parents and progeny of the total polymorphism information content detection (PIC), observed heterozygosity (Ho) and expected heterozygosity (He) were 0.679,0.714 and 0.725. in 95% of the total level identified 203 full sib families, every mother tree with 18 male parent to produce progeny. In the free pollination condition, Schima superba clonal seed orchard of selfing rate was 1.5%, the phenomenon of self is very weak, the mating pattern dominated by outcrossing. The male reproductive contribution rate between 0.49%~7.77%, average 2.44%. as the main pollination seed orchard in Schima superba distance 0~60 m, detect the maximum pollination distance is 120 m, which produced 0~60 m in the range of Pollination Progeny ascertained father population of more than 80%. The book of pollen contamination in seed orchards, that is the comprehensive analysis 7.01%., Schima seed orchard pollen spread evenly, seed orchard and flower powder pollution source isolation The distance should be above 60m and above 120 m to prevent pollen contamination from pollinating insects in long distance pollination activities.

【學(xué)位授予單位】：中國(guó)林業(yè)科學(xué)研究院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S792.99
，

本文編號(hào)：1367060