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作物学报 ›› 2023, Vol. 49 ›› Issue (8): 2133-2143.doi: 10.3724/SP.J.1006.2023.21056

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦矮秆突变体Xu1801的鉴定及其矮化效应分析

苏在兴(), 黄忠勤, 高闰飞, 朱雪成, 王波, 常勇, 李小珊, 丁震乾, 易媛()   

  1. 江苏徐淮地区徐州农业科学研究所, 江苏 徐州 221131
  • 收稿日期:2022-08-21 接受日期:2023-02-10 出版日期:2023-08-12 网络出版日期:2023-03-13
  • 通讯作者: 易媛
  • 作者简介:E-mail: szaixing@163.com
  • 基金资助:
    徐州市重点研发(现代农业)计划项目(CARS-03);财政部和农业农村部国家现代农业产业技术体系建设专项(KC21131)

Identification of wheat dwarf mutant Xu1801 and analysis of its dwarfing effect

SU Zai-Xing(), HUANG Zhong-Qin, GAO Run-Fei, ZHU Xue-Cheng, WANG Bo, CHANG Yong, LI Xiao-Shan, DING Zhen-Qian, YI Yuan()   

  1. Jiangsu Xuhuai Regional Institute of Agricultural Science, Xuzhou 221131, Jiangsu, China
  • Received:2022-08-21 Accepted:2023-02-10 Published:2023-08-12 Published online:2023-03-13
  • Contact: YI Yuan
  • Supported by:
    Key Research and Development Program (Modern Agriculture) of Xuzhou(CARS-03);Key Research and Development Program (Modern Agriculture) of Xuzhou(KC21131)

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摘要：

在徐麦35原种繁育过程中发现1株天然矮秆突变体, 经连续多代单株选择, 该突变体株高性状稳定遗传, 将其命名为Xu1801 (徐1801)。为研究Xu1801矮化原因, 进行了农艺性状调查、倒伏指数分析、苗期赤霉素处理和矮秆相关基因检测。调查发现, Xu1801平均株高68.16 cm, 较野生型徐麦35株高降低24.05%, 其矮化效应表现为节间减少和各节间极显著缩短; 其中, 第5节间效应值最大, 达44.16%。Xu1801的旗叶宽度、叶绿素含量、小穗密度极显著高于徐麦35, 其他农艺性状与徐麦35差异较小。Xu1801产量达8604.17 kg hm-2, 低于徐麦35; 而蛋白质含量、湿面筋、沉降值等品质指标均显著优于徐麦35。Xu1801茎秆粗壮, 各节间充实度较好, 倒伏指数47.73, 极显著低于徐麦35。GA3处理可知, Xu1801和徐麦35同属于赤霉素反应不敏感型种质。分子标记检测发现, Xu1801可能含有Rht-D1b、Rht4、Rht8、Rht9和Rht12等矮秆相关基因; WMC317 (Rht4)和BARC102 (Rht5)在Xu1801和徐麦35之间呈现出不同条带, 其他标记扩增条带在两者之间较为一致。综上, Xu1801株高偏矮、抗倒伏能力强, 产量和品质性状协调, 具有一定的生产应用潜力, 同时携带多个矮秆基因, 可作为小麦株高遗传分析的种质资源。

关键词: 小麦, 矮秆突变, 倒伏指数, 赤霉素, 分子标记

Abstract:

A natural dwarf mutant was discovered in the purification and rejuvenation of Xumai35 (XM35) foundation seeds. After multiple generations of individual selection, the dwarf trait of the mutant named Xu1801 temporarily could be stably inherited. To explore the cause of dwarfing phenotype, agronomic traits, lodging index analysis, gibberellin treatment at seedling stage, and dwarf-related genes detection were carried out. The results showed that the average plant height was 68.16 cm in Xu1801, shorter by24.05% than the wild-type XM35. The dwarfing effect of Xu1801 was manifested in the reduction of internode number and the extremely significant shortening of internode length. Among all internodes, the 5th internode had the largest dwarfing effect (44.16%). Flag leaf width, chlorophyll content, and spikelet density were extremely significantly higher in Xu1801 compared with XM35, and there was no difference in other agronomic traits. Compared with XM35, the grain yield of Xu1801 was lower by 8604.17 kg hm-2, while protein content, wet gluten content, and sedimentation volume of Xu1801 were significantly improved. The stem of Xu1801 was thick, and the internode fullness was extremely significantly higher than that of XM35. The lodging index was 47.73, extremely significantly lower than that of XM35, indicating its outstanding lodging-resistance ability. The detection of dwarfing gene-linked molecular markers revealed that Xu1801 might contain Rht-D1b, Rht4, Rht8, Rht9, and Rht12. Except for WMC317 (Rht4) and BARC102 (Rht5) revealed different amplification bands between Xu1801 and XM35, the amplification bands of other markers were relatively consistent. In conclusion, Xu1801 not only had short plant height and excellent lodging resistance, but also coordinated grain yield and quality characteristics properly, demonstrating the outstanding potential for cultivation. Meanwhile, Xu1801 carried several dwarfing genes and could be utilized as germplasm resources for genetic analysis of wheat plant height.

Key words: wheat, dwarf mutant, lodging index, gibberellin, molecular markers

表1

小麦矮秆基因分子标记及引物序列"

基因
Gene		 引物
Primer name		 引物序列
Primer sequence (5'−3')		 片段长度
Fragment length (bp)		 退火温度
Annealing
temperature (℃)		 染色体
Chromosome		 参考文献
Reference
Rht-B1b
(Rht1)		 BF GGTAGGGAGGCGAGAGGCGAG 237 65 4BS [27]
MR1 CATCCCCATGGCCATCTCGAGCTA
Rht-B1a BF GGTAGGGAGGCGAGAGGCGAG
WR1 CATCCCCATGGCCATCTCGAGCTG
Rht-D1b
(Rht2)		 DF CGCGCAATTATTGGCCAGAGATAG 254 63 4DS
MR2 CCCCATGGCCATCTCGAGCTGCTA
Rht-D1a DF2 GGCAAGCAAAAGCTTCGCG 264
WR2 GGCCATCTCGAGCTGCAC
Rht4 WMC317 F TGCTAGCAATGCTCCGGGTAAC 170 58 2BL [28],[29]
WMC317 R TCACGAAACCTTTTCCTCCTCC
Rht5 BARC102 F GGAGAGGACCTGCTAAAATCGAAGACA 200 58 3BS
BARC102 R GCGTTTACGGATCAGTGTTGGAGA
Rht8 WMC503 F GCAATAGTTCCCGCAAGAAAAG 275 55 2DS
WMC503 R ATCAACTACCTCCAGATCCCGT
Xgwm261 F CTCCCTGTACGCCTAAGGC 192 55
Xgwm261 R CTCGCGCTACTAGCCATTG
Rht9 BARC151 F TGAGGAAAATGTCTCTATAGCATCC 220 55 5AL
BARC151 R CGCATAAACACCTTCGCTCTTCCACTC
Rht12 WMC410 F GGACTTGAAAAGGAAGCTTGTGA 114 61 5AL
WMC410 R CATGGATGGCATGCAGTGT
Rht13 WMS577 F ATGGCATAATTTGGTGAAATTG 130 55 7BS
WMS577 R TGTTTCAAGCCCAACTTCTATT

表2

徐麦35和矮秆突变体Xu1801株高、节间数、节间长度比较"

性状
Trait		 徐麦35
XM35 (cm)		 Xu1801
(cm)		 矮化值
Shorten value (cm)		 矮化效应
Dwarfing effect (%)
株高Plant height 89.74±2.57 68.16±1.60** 21.58 24.05
第1节间长度 1st internode length 27.85±2.26 21.61±1.89** 6.24 22.41
第2节间长度 2nd internode length 16.29±0.77 14.04±0.54** 2.25 13.81
第3节间长度 3rd internode length 12.46±0.78 9.24±0.35** 3.22 25.84
第4节间长度 4th internode length 11.83±0.56 8.10±0.29** 3.74 31.57
第5节间长度 5th internode length 8.90±1.83 4.97±1.96** 3.93 44.16
第6节间长度 6th internode length 3.17±1.51 0.00±0.00 — —

表3

徐麦35和矮秆突变体Xu1801不同农艺性状的比较"

性状Trait 徐麦35 XM35 Xu1801
旗叶长度Flag leaf length (cm) 18.96±1.53 19.48±2.18
旗叶宽度Flag leaf width (cm) 2.28±0.10 2.48±0.13**
旗叶叶面积Flag leaf area (cm2) 32.59±3.64 36.41±5.37
叶绿素含量Leaf chlorophyll content (SPAD) 53.82±0.35 56.82±0.22**
穗长Spike length (cm) 11.04±0.56 10.58±0.41
每穗小穗数Spikelets number per spike 24.40±1.51 24.90±0.57
不育小穗数Empty spikelet number per spike 1.10±0.32 0.90±0.57
小穗密度Spikelet density 2.21±0.08 2.36±0.07**
芒长Awn length (cm) 3.40±0.35 3.19±0.30
籽粒长度Grain length (mm) 5.73±0.09 5.64±0.03
籽粒宽度Grain width (mm) 3.00±0.07 3.12±0.02

图1

徐麦35和Xu1801的主要表型特征 B中I为第1节间(穗下节)、II~VI依次为第2节间至第6节间。"

表4

徐麦35和矮秆突变体Xu1801产量和品质性状的比较"

性状 Trait 徐麦35 XM35 Xu1801
穗数 Spike number (×104 hm-2) 601.45±27.35 523.19±11.16**
穗粒数 Grain number per spike 46.82±0.33 49.61±0.71*
千粒重 1000-grain weight (g) 37.16±0.44 38.60±0.30
产量 Yield (kg hm-2) 9175.60±343.18 8604.17±369.25
容重Volume weight (g L-1) 831.67±4.04 823.33±2.31
蛋白质含量Protein content (%) 9.96±0.33 11.43±0.24*
蛋白质产量Protein yield (kg hm-2) 913.58±30.40 983.17±20.33
湿面筋含量Wet gluten content (%) 21.88±0.53 25.04±0.04*
沉降值Sedimentation volume (mL) 20.67±0.97 24.60±0.44*

表5

徐麦35和矮秆突变体Xu1801各节间充实度"

各节间充实度
The filling degree of internode		 徐麦35茎秆
Stem of XM35 (mg cm-1)		 Xu1801茎秆
Stem of Xu1801 (mg cm-1)
第1节间 1st internode 11.34±0.61 14.62±1.06**
第2节间 2nd internode 20.82±0.74 28.01±1.09**
第3节间 3rd internode 23.92±1.94 31.71±2.63**
第4节间 4th internode 26.88±0.90 33.70±4.52**
第5节间 5th internode 27.99±1.00 35.35±3.25**

图2

Xu1801和徐麦35茎秆倒伏指数分析 **表示在0.01概率水平差异显著。"

图3

外源赤霉素(GA3)对小麦矮秆突变体Xu1801等3个品种(系)苗期胚芽鞘伸长的影响 *和**分别表示10 μmol L-1和100 μmol L-1 GA3条件下在0.05和0.01概率水平与CK存在差异显著。XM35: 徐麦35; SM3: 苏麦3号。"

图4

Rht1和Rht2电泳检测结果 A: Rht-B1a、Rht-B1b (Rht1)基因检测结果; B: Rht-D1a、Rht-D1b (Rht2)基因检测结果。CS: 中国春; XY6: 小偃6号(CK); SN33: 陕农33 (CK); XM35: 徐麦35; SM3: 苏麦3号; HM20: 淮麦20。M: marker, B500331-0250 250 Preps 100~600 bp (Sangon Biotech)。a为Rht1和Rht2野生型; b为Rht1和Rht2突变型。"

图5

小麦矮秆相关基因分子标记检测 M: marker, B500331-0250 250 Preps 100~600 bp (Sangon Biotech)。泳道1~5依次为: Xu1801、徐麦35、苏麦3号、淮麦20、矮抗58。Rht8 (503)、Rht8 (261)分别对应WMC503和Xgwm261标记。"

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