摘要
选用我国主要麦区主栽品种和高代品系 432 份和株高差异大的组合中优 9507 和
CA9632 后代 70 个 DH 株系，用 STS 和 SSR 分子标记检测了矮秆基因 Rht-D1b 和 Rht8 的
分布规律，验证了基于 PCR 的标记在分子标记辅助育种中的可用性，分析了不同矮秆基
因型对株高和小麦产量构成因素的影响。主要结果如下：
1.矮秆基因 Rht-D1b 的分布
利用基于 PCR 的分子标记可以准确检测小麦品种（系）的 Rht-D1b 矮秆基因型。
Rht-D1b 基因全国平均频率为 47.5%，黄淮冬麦区高达 58.7%，北部冬麦区、长江中下
游冬麦区和西南冬麦区分别是 40.9%和 36.8%和 42.5%；北部春麦区为 50%，东北春麦区
最低，为 13.3%，西北春麦区和新疆冬春麦区的分布频率相同，皆为 22.2%。
2.矮秆基因 Rht8 的分布
微卫星 WMS261 标记位点 192bp 的 PCR 扩增片段与 Rht8 紧密连锁，可作为其分子标
记。全国 Rht8 基因分布频率为 45.1%。黄淮冬麦区最高，为 57.7%；西南冬麦区次之，
为 52.9%；北部冬麦区为 34.9%，长江中下游冬麦区分布频率最低，为 15.8%。北部春
麦区和新疆冬春麦区 Rht8 基因的分布频率相当，分别为 25.0%和 22.2%，东北春麦区
和西北春麦区分布频率较低，分别为 6.7%和 11.1%。
3. 标记的可用性验证
432 份小麦材料 Rht-D1b 和 WMS261 标记的分子检测和系谱分析表明，找到 3 个可
以用来检测不同矮秆基因类型的标记。引物 3 针对正常的 Rht-D1a 野生型基因扩增出一
条 264bp 的特异带,引物 4 对正常的 Rht-D1b 突变型基因扩增出一条长度 254bp 的特异
带,引物 5 对 WMS261 标记位点可扩增出 192bp 长度的特异带，其鉴定与 Rht8 基因系谱
分析结果基本一致，Rht-D1b 基因的 PCR 标记也与该基因的鉴定和系谱分析结果基本相
符。通过对这三对引物的重复性验证表明，在系谱关系明确的情况下，微卫星 WMS261
及基于 PCR 的 Rht-D1b 基因的标记可以分别用于对品种 Rht8 基因和 Rht-D1b 基因型的
鉴定以及育种世代该基因型的筛选。
4. 矮秆基因对产量构成因素的影响
矮秆基因具有降低株高的作用，Rht8 属于弱效应降低株高基因，降秆能力 6.0cm
左右，Rht-D1b 属于中度强度的降秆基因，降秆能力在 10.0cm 左右。不同环境条件下，
ii
新疆农业大学硕士学位论文
相同基因型在株高、穗粒数和千粒重三性状的差异均达到 1%显著水平。相同环境条件
下，亲本中优 9507 与基因型 rhtrht、Rht8 和 Rht-D1b+Rht8 的株高性状差异不明显，
变异范围为 88.6—97.5cm，但与 CA9632(父本)和 Rht-D1b 基因差异显著，平均株高分
别是 77.4cm 和 88.6cm。株高主效 QTL 有三个，分别定位在染色体 2DS、4DS 和 6BS 上，
前两个 QTL 分别与 Rht8 基因和 Rht-D1b 基因对应，来自父本 CA9632，对株高有正向降
低作用，定位在 6BS 染色体上的主效 QTL，对应的矮秆基因未知，来源于母本中优 9507，
和上述两个株高 QTL 的作用相反，对株高降低有负向作用。
Totally 432 cultivars and advanced lines from major Chinese wheat regions and 70 DH
lines of Zhongyou 9507 and CA 9632 with significant difference in plant height were
detected by two STS markers and one microsatellite WMS 261 for the semi-dwarfing genes
Rht-D1b and Rht8. The distribution frequencies of the dwarfing genes were determined and
the utilization of the three markers in the validation and effects of dwarfing genes on the
plant height in bread wheat (Triticum aestivum L.) were tested. The results were summarized
below:
1. Frequency of Rht-D1b
Two PCR-based STS markers could be used to test the presence of Rht-D1b in wheat
genotypes.The distribution frequency of Rht-D1b gene in Northern Winter Wheat Zone,
Yellow & Huai River Facultative Winter Wheat Zone, Middle & Low Yangtze Valley Winter
Wheat Zone, Southwestern Winter Wheat Zone, Northern Spring Wheat Zone, Northwestern
Spring Wheat Zone, Northwestern Spring Wheat Zone and Xinjiang Winter-Spring Wheat
Zone was 47.5%., 40.9%, 58.7%, 36.8%, 42.5%, 50.0%, 22.2%, 13.3% and 22.2%,
respectively.
2. Frequency of Rht8 gene
A wheat microsatellite marker, WMS 261, whose 192bp allele closely linked to the
dwarfing gene Rht8 on chromosome 2DS, was used to screen 432 wheat genotypes and 70
DH lines from Zhongyou 9507 and CA 9632 to access the variation at this locus. On
average,the frequency of Rht8 specific to WMS261 marker in China was 45.1%. The highest
frequency of Rht8 was found in Yellow & Huai River Facultative Winter Wheat Zone with
57.7% ,followed by Southwestern Winter Wheat Zone with 52.9%. The frequency of Rht8
gene in Northern winter wheat and Yangtze Valley was 34.9% and 15.8%, respectively. In
Northern Spring Wheat Zone, Northwestern Spring Wheat Zone, Northeastern Spring Wheat
Zone and Xinjiang Winter Spring Wheat Zone, the frequency of Rht8 gene was 25.0%,
i
11.1% , 6.7% and 22.2%, respectively.
3.Validation of PCR marker
The results of molecular detection of 432 genotypes indicated that 3 molecular markers
could be used to detect the semi-dwarfing genes Rht-D1b and Rht8. The primer 3 could
amplify one diagnostic band of 264bp in the wild type gene Rht-D1a, the primer 4 for one
diagnostic band of 254bp in mutant type gene Rht-D1b, and the primer 5 could amplify 4
different diagnostic bands of 165bp, 174bp, 192bp and 202bp for the detection of Rht8. The
results were identical to the ones of pedigree analyses, and the same for the gene Rht-D1b.
Therefore, if the pedigree of a wheat cultivar was clear, the two STS markers and WMS261
could be applied to examine and screen genotypes for semi-dwarfing genes Rht-D1b and
Rht8 in wheat breeding programmes.
4. Effects of dwarfing genes and QTLs on plant height
Rht8 was a weak reducing plant height by about 6.0cm and Rht-D1b was a middle
reduing height gene by about 10.0 cm. A significant difference at 1% level was found in plant
height, spike kernel number and thousand grain weight under different environments,
whereas the difference in plant height between Zhongyou 9507 and the genotypes of Rht8 or
Rht-D1b+Rht8 was not significant with a range of 88.6—97.5cm under the same growing
condition, while a significant difference was detcted between Zhongyou 9507 and CA 9632
or the genotype of Rht-D1b whose plant heights were 77.4cm and 88.6cm. Three major QTLs
were located on chromosomes of 2DS, 4DS and 6BS, repectively and two major QTLs
corresponding to dwarfing genes Rht8 and Rht-D1b derived from CA 9632 showed positive
effect on plant height, one major QTL from Zhongyou 9507 with negetive effect on plant
height.
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