苎麻（Boehmeria nivea（L.）Gaud）高细度种质筛选及其纤维发育相关基因研究
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摘要
苎麻(Boehmeria nivea(L.)Gaud)是一种重要的韧皮纤维作物,其纤维是麻类纤维中优良的纤维,是纺织工业的重要原料,以其独特的优点和用途在国民经济中起着不可缺少和替代的重要作用。而现有苎麻品种的纤维质量难以满足纺织高支、细薄产品的要求。本研究对国家种质长沙苎麻圃中的213份材料进行了鉴定评价,以筛选的纤维细度对比鲜明的种质为材料,检测纤维发育相关基因在不同组织、不同品种、不同时期的表达情况。获得的主要结果如下:
1.单蔸产量与株高、茎粗、皮厚、有效株率和鲜皮出麻率达到极显著正相关。原麻长与株高、茎粗、皮厚、有效株率、鲜皮出麻率和单蔸产量达极显著正相关。纤维细度与株高、茎粗、皮厚、有效株率和鲜皮出麻率达极显著负相关。
2.苎麻单蔸产量和纤维细度存在丰富的遗传变异,单蔸产量的最大变异系数47.27%,最小值为0.08%。纤维细度的最大变异系数29.56%,最小值0.07%。
3.筛选出高产种质(单蔸产量≥110g)56份,优质(纤维细度≥2200m/g)种质38份,高产(单蔸产量≥110g)优质(纤维细度≥2200 m/g)的种质5份。
4.筛选出能同时稳定扩增目的基因FB27和CFE-1片段的最优化反应条件:最佳镁离子浓度为3.0 mmol L-1,退火温度为55.6℃,35个循环。
5.在根中纤维蛋白FB27基因的相对表达量为0.294,茎皮为0.845,叶为0.347,茎尖为0.267,茎骨为0.719,总的规律是茎皮>茎骨>叶>根>茎尖。在各器官均有表达,FB27基因无组织表达特异性,但在苎麻纤维生长增粗期的茎部表达量最高,尤其是茎皮。在高纤维细度种质中相对表达量较高,在低纤维细度种质中相对表达量较低;在不同的生长季FB27基因的相对表达量为:头麻>三麻>二麻。在同季麻不同生长发育时期,伸长增粗期FB27基因的相对表达量较高,苗期和工艺成熟期较低。
6.纤维表达蛋白CFE-1基因在各器官均有表达,其相对表达量总的规律是茎皮>茎骨>茎尖>叶>根。在高纤维细度种质中相对表达量较高,显著高于在低纤维细度种质中相对表达量;在不同的生长季CFE-1基因的相对表达量为,头麻>三麻>二麻。在同季麻不同生长发育时期,伸长增粗期CFE-1基因的相对表达量较高,苗期、工艺成熟期较低。
Ramie (Boehmeria nivea (L.) Gaud)is an important bast fiber crop, whose fiber which is one of the best bast fibers, is widely used in textile industry and plays an indispensable and irreplaceable role in the national economy. But now the fiber quality of ramie accessions are difficult to meet the high-textiles, small thin product requirements. Agronomic characters and bast fibre quality characters of totally 213 collected accessions of ramie were evaluated, and the genes expression related with fiber development were detected in different development period, between different genotypes by the semi-quantitative PCR. The main results were as follow:
1. Yeild per rootstock and plant Height , diameter of stem, thickness of bark, rate of effective tiller, fibre rate of fresh bark existed a significant positive correlation relationships. There was a significant positive correlation between the raw ramie length and plant Height, diameter of stem, thickness of bark, rate of effective tiller, fibre rate of fresh bark, yeild per rootstock .It was a significant negative correlation between fibre fineness and plant Height, diameter of stem, thickness of bark, rate of effective tiller, fibre rate of fresh bark.
2. There was significantly variance among agronomical characters and bast fibre quality characters. The Maximum coefficient of variation of yeild per rootstock was 47.27 %, the minimum is 0.08%, and that The Maximum coefficient of variation of fibre fineness was 29.56% and the minimum was 0.07%.
3. 56 varieties were found to be high yield(≥110g). 38 varieties were identified as germplasm with good fineness(≥2200m/g). 5 varieties were identified as the accessions with excellently high yield (≥110g)and high fineness(≥2200m/g).
4. The best optimal conditions amplifying fragments FB27 and CFE-1 at the same time could be screened: the best concentration of magnesiumions is 3.0 mmol L-1, annealing temperature is 55.6℃, 35 cycles.
5. The relative expression of FB27 gene is 0.294 in the root, 0.845 in skin stem, 0.347 in leaves, 0.267 in shoot tip, 0.719 in stem bone. The general rule is that, bast> stem bone>leaf> root>shoot tip. FB27 expressed in various organs showed that FB27 gene expression is non-specific organizations, but the highest expression in the term of strong growth of stem, in particular, bast. The relative higher expression existed in high-fiber fineness germplasms, and the relative lower expression existed in low-fiber fineness germplasms. The relative gene expression of FB27 is that first-crop> third-crop > second-crop.
6. CFE-1 gene was expressed in various organs, the general rule of the relative expression was that, bast> stem bone> shoot tip> leaf> root. The relatively higher expression existed in high-fiber fineness germplasms was significantly higher than those in low-fiber fineness germplasms. The relative gene expression of CFE-1 in Harvest periods was that first-crop> third -crop > second -crop.
1.单蔸产量与株高、茎粗、皮厚、有效株率和鲜皮出麻率达到极显著正相关。原麻长与株高、茎粗、皮厚、有效株率、鲜皮出麻率和单蔸产量达极显著正相关。纤维细度与株高、茎粗、皮厚、有效株率和鲜皮出麻率达极显著负相关。
2.苎麻单蔸产量和纤维细度存在丰富的遗传变异,单蔸产量的最大变异系数47.27%,最小值为0.08%。纤维细度的最大变异系数29.56%,最小值0.07%。
3.筛选出高产种质(单蔸产量≥110g)56份,优质(纤维细度≥2200m/g)种质38份,高产(单蔸产量≥110g)优质(纤维细度≥2200 m/g)的种质5份。
4.筛选出能同时稳定扩增目的基因FB27和CFE-1片段的最优化反应条件:最佳镁离子浓度为3.0 mmol L-1,退火温度为55.6℃,35个循环。
5.在根中纤维蛋白FB27基因的相对表达量为0.294,茎皮为0.845,叶为0.347,茎尖为0.267,茎骨为0.719,总的规律是茎皮>茎骨>叶>根>茎尖。在各器官均有表达,FB27基因无组织表达特异性,但在苎麻纤维生长增粗期的茎部表达量最高,尤其是茎皮。在高纤维细度种质中相对表达量较高,在低纤维细度种质中相对表达量较低;在不同的生长季FB27基因的相对表达量为:头麻>三麻>二麻。在同季麻不同生长发育时期,伸长增粗期FB27基因的相对表达量较高,苗期和工艺成熟期较低。
6.纤维表达蛋白CFE-1基因在各器官均有表达,其相对表达量总的规律是茎皮>茎骨>茎尖>叶>根。在高纤维细度种质中相对表达量较高,显著高于在低纤维细度种质中相对表达量;在不同的生长季CFE-1基因的相对表达量为,头麻>三麻>二麻。在同季麻不同生长发育时期,伸长增粗期CFE-1基因的相对表达量较高,苗期、工艺成熟期较低。
Ramie (Boehmeria nivea (L.) Gaud)is an important bast fiber crop, whose fiber which is one of the best bast fibers, is widely used in textile industry and plays an indispensable and irreplaceable role in the national economy. But now the fiber quality of ramie accessions are difficult to meet the high-textiles, small thin product requirements. Agronomic characters and bast fibre quality characters of totally 213 collected accessions of ramie were evaluated, and the genes expression related with fiber development were detected in different development period, between different genotypes by the semi-quantitative PCR. The main results were as follow:
1. Yeild per rootstock and plant Height , diameter of stem, thickness of bark, rate of effective tiller, fibre rate of fresh bark existed a significant positive correlation relationships. There was a significant positive correlation between the raw ramie length and plant Height, diameter of stem, thickness of bark, rate of effective tiller, fibre rate of fresh bark, yeild per rootstock .It was a significant negative correlation between fibre fineness and plant Height, diameter of stem, thickness of bark, rate of effective tiller, fibre rate of fresh bark.
2. There was significantly variance among agronomical characters and bast fibre quality characters. The Maximum coefficient of variation of yeild per rootstock was 47.27 %, the minimum is 0.08%, and that The Maximum coefficient of variation of fibre fineness was 29.56% and the minimum was 0.07%.
3. 56 varieties were found to be high yield(≥110g). 38 varieties were identified as germplasm with good fineness(≥2200m/g). 5 varieties were identified as the accessions with excellently high yield (≥110g)and high fineness(≥2200m/g).
4. The best optimal conditions amplifying fragments FB27 and CFE-1 at the same time could be screened: the best concentration of magnesiumions is 3.0 mmol L-1, annealing temperature is 55.6℃, 35 cycles.
5. The relative expression of FB27 gene is 0.294 in the root, 0.845 in skin stem, 0.347 in leaves, 0.267 in shoot tip, 0.719 in stem bone. The general rule is that, bast> stem bone>leaf> root>shoot tip. FB27 expressed in various organs showed that FB27 gene expression is non-specific organizations, but the highest expression in the term of strong growth of stem, in particular, bast. The relative higher expression existed in high-fiber fineness germplasms, and the relative lower expression existed in low-fiber fineness germplasms. The relative gene expression of FB27 is that first-crop> third-crop > second-crop.
6. CFE-1 gene was expressed in various organs, the general rule of the relative expression was that, bast> stem bone> shoot tip> leaf> root. The relatively higher expression existed in high-fiber fineness germplasms was significantly higher than those in low-fiber fineness germplasms. The relative gene expression of CFE-1 in Harvest periods was that first-crop> third -crop > second -crop.
引文
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