【作者】 栾启福；

【导师】 张守攻；

【作者基本信息】 中国林业科学研究院 ， 林木遗传育种， 2010， 博士

【摘要】 本研究以南方湿地松、火炬松为母本,以加勒比松三个种(变种)为父本,开展了杂交试验、杂种分子鉴别和杂种生长、干形和基本密度的配合力分析及早期选择研究。为了减少试验分析误差,试验中以单株(个)数据为单元,充分利用原始数据信息,利用SAS软件和ASReml软件线性模型,开发出以松树EST序列为基础的功能基因标记,用于杂种鉴别和优良单株筛选,主要研究及研究结果如下:一、以母本湿地松和火炬松半同胞家系测定为基础,以遗传力、遗传增益、配合力和育种值为指标,对母本遗传背景进行分析,筛选出优良家系作为杂交选育首选的亲本材料。研究结果显示,湿地松和火炬松半同胞家系生长性状的家系遗传力在0.3000~0.6000之间,为中等-偏低遗传水平,而单株遗传力在0.1000-0.3000之间,其中湿地松树高的单株遗传力较高;而遗传增益分析进一步显示,火炬松家系选择比湿地松家系选择效果要好。二、以火炬松×加勒比松F1代群体为研究对象,从数十万松树PGI数据中筛选出13个与赤霉酸代谢有关的EST序列,并设计了这13条序列的EST引物对,从中筛选出4对引物作为F1代检测的较好的标记。4对引物PCR分析显示,在2个亲本和39个子代中共扩增出1170个多态性位点,其中杂种F1代扩增出的位点数中有54.79%与父本相同,52.31%与母本相同,有24.79%的特异性位点既不属于父本、也不属于母本;4个引物检测的30个等位基因位点中有7个与苗高有显著或极显著相关,利用这7个位点联合选择可以选择到生长表现较优的单株,为早期选择提供了较好的分子标记。由此表明,母本(火炬松)和父本(加勒比松)杂交能够获得双亲遗传物质新的杂种,同时可能产生染色体的特异性变异。三、以杂种球果、种子和幼苗为研究对象,研究其变异特性。研究结果表明,火炬松×加勒比松以及湿地松×加勒比松杂种的千粒重平均值均比对应的母本轻。而一年生苗高平均值显示,杂种苗高均高于对应母本,显示杂种苗期生长优势。杂种及其对应母本的方差、标准差、极差、变异系数及平均值显示杂种的所有性状的离散参数均比对应母本大,由此说明杂交可创造出比母本更大的遗传变异,为遗传选择提供了更大空间。四、以测交模型为主,研究了湿地松×加勒比松、火炬松×加勒比松部分生长和材性指标的遗传力和配合力。湿地松×加勒比松配合力分析显示,母本湿地松的一般配合力效应显著;遗传力分析显示,母本湿地松基本密度和干形遗传力较高,而父本加勒比松生长量的遗传力较高。表型和遗传相关分析显示,湿地松×加勒比松杂种基本密度与生长呈极显著负相关,其遗传负相关也很大,不利于基本密度和生长性状的同步改良,而基本密度与通直度和分枝大小可同步改良。火炬松×加勒比松研究显示,生长性状母本单株遗传力与父本单株遗传力处于中、低水平。基本密度主要表现为母本遗传,而通直度表现为父本遗传力大于母本遗传力。性状相关分析表明,生长性状和基本密度之间存在较高的遗传负相关和显著的表型负相关,同步改良有难度;而生长性状和通直度以及侧枝大小之间可以同步改良,只是侧枝大小受环境影响较大,可以通过栽培措施获得同步改良效果。侧枝和通直度表型之间表现为正相关,可以同步改良。亲本配合力高低与亲本的来源及变种属性有关,选择优良亲本可以增加获得更高的杂种特殊配合力的几率。基本密度由于特殊配合力效应显著,因此虽然和生长性状负相关,可以利用特殊配合力选择生长和基本密度均较好的杂交组合,但几率较低。五、研究得出Pilodyn方法评估杂交松基本密度的回归方程。研究显示,湿地松×加勒比松Pilodyn测定值与基本密度之间呈显著负相关,由此说明,利用Pilodyn探测值估测湿地松×加勒比松家系可行,为便捷测量基本密度提供了依据;利用同样的方法建立了火炬松×加勒比松Pilodyn测定值与基本密度之间的回归方程,回归关系极显著。由此可知,利用Pilodyn探测值估测火炬松×加勒比松家系同样便捷可行。六、研究显示可以通过早期选择获得较高的遗传增益,选择年份越高,选择效果越好,但是没有到决选树龄。火炬松×加勒比松材积的年度间遗传相关较显著,从实验中遗传相关系数和表型相关系数得知,材积选择有效,但也不到决选树龄。通过单株模型对火炬松×加勒比松杂种及其母本半同胞家系多年多点试验综合分析显示,其杂交组合的树高和地径的特殊配合力效应值较高。父本加勒比松洪都拉斯变种整体表现比巴哈马变种好,并且在树高和地径两个性状上表现比较一致。母本火炬松沿海的优选种源无性系一般配合力较好,与加勒比松杂交时的特殊配合力也较好,根据一般配合力和特殊配合力值可以为营建种子园以及其它生产群体提供科学依据。更多还原

【Abstract】 Interspecific hybrid test、hybrid identification by moleculer markers and estimation of heritability and combining ability were done with female parents Pinus elliottii Engelman var. elliottii×P. caribaea Morelet(PEE)and P. taeda L.(PTA) and male parents P. caribaea Morelet(PCA, including PCC(P. caribaea Morelet var. caribaea ), PCH( Pinus caribaea var. hondurensis Barrett et Golfari) and PCB (Pinus caribaea var. bahamensis Barrett et Golfari)) using general linear model(GLM) or mixed linear model of software SAS and ASReml and EST-SSR markers. Main results were as follows:(1)Genetic background of the female parents was analyzed with half-sib families of PEE and PTA using the evaluating parameters of genetic ability、combing ability、genetic gain and breeding value。The results showed that mean heritability of growth traits for half-sib families of PEE and PTA was 0.3-0.6, which was medium-low level;while the heritability for single trees ( h s2)was 0.1-0.3。Better selection effects was got by the stem height of PEE than PTA because of higher h s2 for PEE,and family selection was the better method for PTA。(2)Molecular markers reletated to GAs metabolism were developed from thousands of EST sequences in PGI data. 1170 gene-locus were amplified in 2 parents and their 39 F1 hybrids with 4 primer-pairs. The hybrids had 54.79% of the 1170 locus same as the male parent and 52.31% same as the female parent, and 24.79% was different from the parents, which showed the hybrid experiments made the new genetic genotype . 7 allele locus seleted from all 30 allele locus amplified by 4 primer pairs were related significantly to the traits of seedling height, and better seedlings in growth traits can be selected with higher positive rate using the 7 locus, which were the good molecular markers for early selection.(3) Genetic variation for the quality and size of cones and seeds of different hybrids and their parents were studied with T-test。The results showed that: length and width of single cone of Pinus taeda L.×P. caribaea Morelet(PTA×PC) were little smaller than the size of their female parents, while the quality of single cone(39.00g) of the hybrids was smaller significantly(P=0.05) than that(48.00g) of their female parents. The dispersion degree(Variance, standard deviation, range and CV ) of PEE×PCA and PTA×PCA were greater than their female parents,which showed the hybrids created more variation for selection.(4)Regression equation between the value(x) measured by Piloyn and wood basic density(y) measured by increment borer was constructed, which for PEE×PCA is y = -0.0127x + 0.5638 (P=0.0021,Coefficient of determination R2 = 0.3685);the equation for PTA×PCA is y = -0.0071x + 0.4668(P<0.0001,R2 = 0.4595)。(5)Heritability and combing ability for part of the growth and quality traits of PEE×PCA and PTA×PCA were estimated by testcross desingn(NCII ). The results showed that the effect on general combing ability(GCA)for the female parent of PEE×PCA was significant and the selection of female parent was important; h s2 of basic density and stem quality for female single trees was 0.2261-0.6382, and h s2 of growth traits for male single trees was 0.1289-0.3248. Genetic correlation and phenotypic correlation between growth traits and basic density of PEE×PCA was negative significantly, and the coefficient of genetic correlation was 1.14-1.16. The correlation between growth traits and stem quality was positive.The analysis of heritability(h2) of PTA×PCA showed that family mean heritability of growth,stem-straightness and wood density for female parents was higher than that for male parents, while the corresponding mean h2 for full-sib families was in the middle of the parents’heritability. The h2 of growth for female parents was 0.2923~0.4699, that for male parents was 0.1919~0.4033. The family mean and single tree h2 of wood basic density for female parents was 0.1764 and 0.0346, respectively, and the single tree h2 of stem-straightness for male and female parents was 0.1627 and 0.0983, respectively. The analysis of genetic correlation showed that the correlation between growth traits and stem-form was positive (|rG| =0.4672~0.3516), and that between growth traits and wood basic density was negative (|rG| =0.2203~0.6798). The ANOVA of combing ability showed that the GCA (general combining ability) effect of male or female parents for growth traits and stem-straightness has significant difference, and their effect of SCA(special combing ability) of combination for wood basic density was significant. The effective value of combing ability for GCA and SCA was estimated and the better parents and hybrids were evaluated. The correlation between genetic background of the parents and genetic differences of the progenies was discussed.(6)Early selection for growth traits for hybrid pines was studied. Genetic correlation coefficient of stem height of PEE×PCA between year 2001 and 2003 was 1.3839, that between year 2003 and 2005 was 2.0552, year 2005 and 2007 was 2.4196, which showed early selction for height was effective. Genetic correlation coefficient of volume of PTA×PCA among year 2003-2005 was 0.732 and 1.0081, and phenotypic correlation coefficient was 0.3927 and 0.8919 (P<0.01).Heritability and combing ability(or breeding value, BV)of PTA×PCA and PTA half-sib families were estimated fully using multi-sites and multi-years data by individual tree model of ASReml software. The results showed that h2 of tree height and diameter at floor height(DFH)for single tree were 0.1811±0.049 and 0.0939±0.0356,respectively.The value of SCA(Special combing ability) of height and DFH for 31-041×26 and 32-031×22 was relatively high. GCA of male parent PCH was totally higher than that of PCB and PCC。Better female parent PTAs were good provenances along the coast of USA such as 31161、31041 and 31062. Individual BV could be used for forward selection for establishing breeding population and parents with best BV plus higher SCA can be used for selection for deployment population (such as new seed orchards). Female parents of elite hybrids PTA×PCA were mainly common provenances(vs cold-resistant provenances), which were from Mississippi, South Carolina and North Carolina, on the selection of seven-year old hybrids planted in Hangzhou , Zhejiang(30°27′N,119°48′E).更多还原