虉草遗传连锁图谱构建与种质资源评价
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摘要
虉草(Phalaris arundinacea L.)作为一种多年生冷季型禾草,以其丰富的地上生物量,高木质纤维素含量和高燃烧值等优势跻身草本能源植物之列。近年来,育种工作者们开始以培育具有良好能源品质性状的目标开展着虉草的育种工作。由于虉草遗传育种工具,例如遗传图谱等的缺乏,以及对现有虉草种质资源能源品质性状了解的欠缺,本研究从构建虉草遗传连锁图谱和虉草种质资源能源品质性状的分析两部分着手,从而协助推进虉草能源植物新品种的育种工作。
本研究第一部分,采用木质素含量有显著差异的虉草双亲和阿魏酸含量有显著差异且高度杂合的双亲,采用“双拟测交”法构建2个作图群体。来自高羊茅和黑麦草的EST-SSR标记、TRAP标记和AFLP标记用于构建虉草遗传连锁图谱。具体结果如下：
1)群体5145中,父本(RC-45)遗传连锁图中共包含有20个连锁群,65个标记共覆盖基因组长度为675.16cM,平均标记密度为10.55cM,基因组估计长度为1286.41cM,图谱覆盖率为52.5%。母本(RC-51)遗传连锁图谱由20个连锁群构成,69个标记共覆盖基因组长度为711.26cM,平均标记密度为10.46cM,基因组估计长度为1226.57cM,图谱覆盖率为58.0%。
2)群体3242中,父本(RC-42)连锁图谱所构建的13个连锁群,75个标记共覆盖基因组长度为421.21cM,平均标记密度为5.69cM。母本(RC-32)连锁图谱所构建的16个连锁群,75个标记共覆盖基因组长度为661.76cM,平均标记密度为8.94cM。
本研究第二部分,针对虉草5个能源品质性状(酸性洗涤纤维[ADF]、中性洗涤纤维[NDF]、酸性洗涤木质素[ADL]、灰分和热值)建立NIRS定标方程,并对来自美国的8个虉草品种和来自6个州的72个虉草材料的5个能源品质性状进行分析,研究结果如下：
1)通过对不同种质材料在两地、两年和两个生长期的大量样本分析获得了虉草ADF、 NDF、ADL、灰分和热值的NIRS定标方程,可以运用到虉草能源品质性状评价及标准制定中。
2)虉草的5个能源品质性状分别于2006和2007年,在美国康奈尔州伊萨卡市(ITH)和威斯康辛州阿灵顿市(ARL)表现出一定程度的差异。已育成的部分虉草品种和部分材料材料具备良好的能源品质。对基因型和环境互作效应的研究发现,5个性状均不受环境影响,ADF、 NDF、ADL和热值受基因型调控,而灰分含量不会受到基因型的调控。
3)ADF与NDF、ADF与ADL、NDF与ADL、NDF与热值之间分别呈现显著正相关关系,而灰分与ADL、灰分与热值之间则呈现显著性负相关关系。五个能源品质性状在2006年与2007年间呈现了显著性正相关关系,在ITH和ARL两地之间也呈现了显著性正相关关系。
4)"Marathon"和“RC-135”可作为生产液态燃料虉草品种选育的基础材料："Chieft、 Palaton、RC-139、RC-140、RC-145"(热值高)和" RC-141、RC-143、RC-181、RC-185、 RC-193、RC-195"(木质素含量高且灰分含量低)可作为生产固态燃料虉草品种选育的基础材料。
Reed canarygrass is a perennial cool-season grass. Due to its high yield, high lignocelluloses content and high calorific value, reed canarygrass became to one of the highest potential bioenergy plants. Recently, breeders worked on breeding reed canarygrass cultivars with good bioenergy quality. The reed canarygrass breeding work moved slowly, since lacking of the genetic tools, such as genetic linkage maps, and understanding less about the bioenergy traits of reed canarygrass gerplasm. To assist the breeding work of reed canarygrass, this study focused on constructing genetic linkage maps of reed canarygrass and analyzing the bioenergy traits of its gerplasm.
In the first study, the two mapping populations were both first generation (F1) which derived from four phenotypically (two for lignin content, another two for ferulate content) divergent, highly heterozygous reed canarygrass phenotypes by "two-way pseudo cross" strategy. The EST-SSR markers from tall fescue and ryegrass, TRAP markers and AFLP markers were used for constructing the genetic linkage maps. The information of the genetic linkage maps as follows,
1) In population5145, the paternal (RC-45) genetic map contained65markers over20linkage groups (LGs). The total map distance of the paternal map was675.16cM, and the mean map distance between markers was10.55cM. The estimated genome size was1286.41cM, and the coverage of the map was52.5%. The maternal (RC-51) genetic map was composed of69markers over20linkage groups (LGs). The total map distance of the male map was711.26cM, and the mean map distance between markers was10.46cM. The estimated genome size was1226.57cM, and the coverage of the map was58.0%.
2) In population5145, the paternal (RC-42) genetic map contained75markers over13linkage groups (LGs). The total map distance of the paternal map was421.21cM, and the mean map distance between markers was5.69cM. The maternal (RC-32) genetic map was composed of75markers over16linkage groups (LGs). The total map distance of the male map was661.76cM, and the mean map distance between markers was8.94cM.
In the second study, the calibration equations of5bioenergy traits (ADF, NDF, ADL, ash and calorific value) were established, which were used to evaluate the bioenergy traits of8reed canarygrass cultivars and72reed canarygrass accessions were collected from6states of U.S. The study results as follows.
1) The reed canarygrass samples collected from two locations, two years and two stages were used to establish the calibration equations of ADF, NDF, ADL, ash and calorific value. The calibration equations can be used for evaluating bioenergy traits of reed canarygrass and setting the standard of the bioenergy traits.
2) The5phenotypic traits of reed canarygrass showed the variation over two years (2006and2007) in Ithaca, NY, USA (ITH) and Arlington, WI, USA (ARL). Although the cultivars had shown the good bioenergy quality, some of the accessions also had potential to become high quality biomass. The genotype and environment interaction showed that the phenotypic traits were not affected by environment, and the genotype affected the bioenergy traits except ash.
3) ADF and NDF, ADF and ADL, NDF and ADL, NDF and calorific value were all positively correlated with each other. The negative correlation was between ash and ADL, ash and calorific value. The5bioenergy traits were all positively correlated between2006and2007. Also, the5bioenergy traits were all positively correlated between ITH and ARL.
4)"Marathon" and "RC-135" can be used as the breeding materials for liquid bioenergy production."Chieft, Palaton, RC-139, RC-140, RC-145"(high calorific value) and "RC-141, RC-143, RC-181, RC-185, RC-193, RC-195"(high lignin content and low ash content) can be used as the breeding materials for solid bioenergy production.
本研究第一部分,采用木质素含量有显著差异的虉草双亲和阿魏酸含量有显著差异且高度杂合的双亲,采用“双拟测交”法构建2个作图群体。来自高羊茅和黑麦草的EST-SSR标记、TRAP标记和AFLP标记用于构建虉草遗传连锁图谱。具体结果如下：
1)群体5145中,父本(RC-45)遗传连锁图中共包含有20个连锁群,65个标记共覆盖基因组长度为675.16cM,平均标记密度为10.55cM,基因组估计长度为1286.41cM,图谱覆盖率为52.5%。母本(RC-51)遗传连锁图谱由20个连锁群构成,69个标记共覆盖基因组长度为711.26cM,平均标记密度为10.46cM,基因组估计长度为1226.57cM,图谱覆盖率为58.0%。
2)群体3242中,父本(RC-42)连锁图谱所构建的13个连锁群,75个标记共覆盖基因组长度为421.21cM,平均标记密度为5.69cM。母本(RC-32)连锁图谱所构建的16个连锁群,75个标记共覆盖基因组长度为661.76cM,平均标记密度为8.94cM。
本研究第二部分,针对虉草5个能源品质性状(酸性洗涤纤维[ADF]、中性洗涤纤维[NDF]、酸性洗涤木质素[ADL]、灰分和热值)建立NIRS定标方程,并对来自美国的8个虉草品种和来自6个州的72个虉草材料的5个能源品质性状进行分析,研究结果如下：
1)通过对不同种质材料在两地、两年和两个生长期的大量样本分析获得了虉草ADF、 NDF、ADL、灰分和热值的NIRS定标方程,可以运用到虉草能源品质性状评价及标准制定中。
2)虉草的5个能源品质性状分别于2006和2007年,在美国康奈尔州伊萨卡市(ITH)和威斯康辛州阿灵顿市(ARL)表现出一定程度的差异。已育成的部分虉草品种和部分材料材料具备良好的能源品质。对基因型和环境互作效应的研究发现,5个性状均不受环境影响,ADF、 NDF、ADL和热值受基因型调控,而灰分含量不会受到基因型的调控。
3)ADF与NDF、ADF与ADL、NDF与ADL、NDF与热值之间分别呈现显著正相关关系,而灰分与ADL、灰分与热值之间则呈现显著性负相关关系。五个能源品质性状在2006年与2007年间呈现了显著性正相关关系,在ITH和ARL两地之间也呈现了显著性正相关关系。
4)"Marathon"和“RC-135”可作为生产液态燃料虉草品种选育的基础材料："Chieft、 Palaton、RC-139、RC-140、RC-145"(热值高)和" RC-141、RC-143、RC-181、RC-185、 RC-193、RC-195"(木质素含量高且灰分含量低)可作为生产固态燃料虉草品种选育的基础材料。
Reed canarygrass is a perennial cool-season grass. Due to its high yield, high lignocelluloses content and high calorific value, reed canarygrass became to one of the highest potential bioenergy plants. Recently, breeders worked on breeding reed canarygrass cultivars with good bioenergy quality. The reed canarygrass breeding work moved slowly, since lacking of the genetic tools, such as genetic linkage maps, and understanding less about the bioenergy traits of reed canarygrass gerplasm. To assist the breeding work of reed canarygrass, this study focused on constructing genetic linkage maps of reed canarygrass and analyzing the bioenergy traits of its gerplasm.
In the first study, the two mapping populations were both first generation (F1) which derived from four phenotypically (two for lignin content, another two for ferulate content) divergent, highly heterozygous reed canarygrass phenotypes by "two-way pseudo cross" strategy. The EST-SSR markers from tall fescue and ryegrass, TRAP markers and AFLP markers were used for constructing the genetic linkage maps. The information of the genetic linkage maps as follows,
1) In population5145, the paternal (RC-45) genetic map contained65markers over20linkage groups (LGs). The total map distance of the paternal map was675.16cM, and the mean map distance between markers was10.55cM. The estimated genome size was1286.41cM, and the coverage of the map was52.5%. The maternal (RC-51) genetic map was composed of69markers over20linkage groups (LGs). The total map distance of the male map was711.26cM, and the mean map distance between markers was10.46cM. The estimated genome size was1226.57cM, and the coverage of the map was58.0%.
2) In population5145, the paternal (RC-42) genetic map contained75markers over13linkage groups (LGs). The total map distance of the paternal map was421.21cM, and the mean map distance between markers was5.69cM. The maternal (RC-32) genetic map was composed of75markers over16linkage groups (LGs). The total map distance of the male map was661.76cM, and the mean map distance between markers was8.94cM.
In the second study, the calibration equations of5bioenergy traits (ADF, NDF, ADL, ash and calorific value) were established, which were used to evaluate the bioenergy traits of8reed canarygrass cultivars and72reed canarygrass accessions were collected from6states of U.S. The study results as follows.
1) The reed canarygrass samples collected from two locations, two years and two stages were used to establish the calibration equations of ADF, NDF, ADL, ash and calorific value. The calibration equations can be used for evaluating bioenergy traits of reed canarygrass and setting the standard of the bioenergy traits.
2) The5phenotypic traits of reed canarygrass showed the variation over two years (2006and2007) in Ithaca, NY, USA (ITH) and Arlington, WI, USA (ARL). Although the cultivars had shown the good bioenergy quality, some of the accessions also had potential to become high quality biomass. The genotype and environment interaction showed that the phenotypic traits were not affected by environment, and the genotype affected the bioenergy traits except ash.
3) ADF and NDF, ADF and ADL, NDF and ADL, NDF and calorific value were all positively correlated with each other. The negative correlation was between ash and ADL, ash and calorific value. The5bioenergy traits were all positively correlated between2006and2007. Also, the5bioenergy traits were all positively correlated between ITH and ARL.
4)"Marathon" and "RC-135" can be used as the breeding materials for liquid bioenergy production."Chieft, Palaton, RC-139, RC-140, RC-145"(high calorific value) and "RC-141, RC-143, RC-181, RC-185, RC-193, RC-195"(high lignin content and low ash content) can be used as the breeding materials for solid bioenergy production.
引文
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