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作物学报 ›› 2021, Vol. 47 ›› Issue (11): 2111-2120.doi: 10.3724/SP.J.1006.2021.04251

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

转PSAG12-IPT基因对棉花叶片衰老及产量和纤维品质的影响

王晔1,2**(), 刘钊2,**, 肖爽2, 李芳军2, 吴霞3, 王保民2, 田晓莉2,*()   

  1. 1北京农学院植物科学技术学院, 北京 102206
    2中国农业大学农学院 / 植物生长调节剂教育部工程研究中心, 北京 100193
    3山西农业大学 / 山西农业科学院棉花研究所, 山西运城 044000
  • 收稿日期:2020-11-21 接受日期:2021-04-26 出版日期:2021-11-12 网络出版日期:2021-06-09
  • 通讯作者: 田晓莉
  • 作者简介:王晔, E-mail: wangyebua@126.com第一联系人：**同等贡献
  • 基金资助:
    国家转基因生物新品种培育重大专项(2011ZX08005-004-008);国家自然科学基金项目(31701337)

Effects of PSAG12-IPT gene expression on leaf senescence, yield, and fiber quality in cotton

WANG Ye1,2**(), LIU Zhao2,**, XIAO Shuang2, LI Fang-Jun2, WU Xia3, WANG Bao-Min2, TIAN Xiao-Li2,*()   

  1. 1College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, China
    2College of Agronomy and Biotechnology, China Agricultural University / China Engineering Research Center of Plant Growth Regulator, Ministry of Education, Beijing 100193, China
    3Shanxi Agricultural University / Cotton Research Institute Shanxi Academy of Agricultural Sciences, Yuncheng 044000, Shanxi, China
  • Received:2020-11-21 Accepted:2021-04-26 Published:2021-11-12 Published online:2021-06-09
  • Contact: TIAN Xiao-Li
  • About author:First author contact:** Contributed equally to this work
  • Supported by:
    National Major Project for Developing New GM Crops(2011ZX08005-004-008);National Natural Science Foundation of China(31701337)

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摘要：

探讨PSAG12-IPT基因导入对棉花叶片衰老性状、产量和纤维品质的影响。本研究以9个转化事件的纯合阳性株系为试验材料, 于2011—2012年在中国农业大学上庄实验站开展研究。结果表明, 转基因株系叶片衰老阶段IPT基因表达量大幅上调, Z+ZR类细胞分裂素含量显著高于野生型冀合321。根据Z+ZR类细胞分裂素含量将9个转PSAG12-IPT基因株系聚类分为抗衰老能力强、弱和中间型3大类, 不同类型株系在叶片衰老阶段的叶绿素和可溶性蛋白含量较野生型增加幅度不同。转基因株系单株成铃数(直径≥2 cm)和铃重与野生型相当或高于野生型, 但各株系第一次收获时的吐絮率显著低于野生型, 且降低幅度与抗衰老能力成正比。这导致抗衰老能力较强和中等的PSAG12-IPT过表达株系(OE-37和OE-38)因有效吐絮铃少而减产, 而抗衰老能力弱的株系(OE-30)籽棉产量与野生型相当。PSAG12-IPT基因过表达株系的纤维长度整齐度指数显著高于野生型, 马克隆值有增加趋势, 其他品质指标与野生型差别不大。

关键词: 棉花, PSAG12-IPT基因, 细胞分裂素, 产量, 纤维品质

Abstract:

A chimeric gene of bacterial IPT and the senescence-specific SAG12 promoter from Arabidopsis (PSAG12-IPT) was overexpressed in cotton, and nine independent transgenic lines were obtained. To identify the leaf senescence characteristics, yield, and fiber quality of PSAG12-IPT cotton plants, we performed the experiments at the Shangzhuang Experimental Station of China Agricultural University in 2011 and 2012. The results showed that the expression of IPT genes in transgenic lines was dramatically up-regulated at leaf aging stage, and the content of Z+ZR-type cytokinin was significantly higher than that of wild type Jihe 321. According to the content of Z+ZR-type cytokinin, nine transgenic lines were clustered into three categories with strong, weak, and moderate anti-aging ability. During leaf senescence period, the contents of chlorophyll and soluble proteins in transgenic lines were positively related to the content of Z+ZR-type cytokinin. Compared to wild type, transgenic lines had a higher or equal number of bolls (diameter ≥ 2 cm) per plant and boll weigh, but their boll-opening rate at the first harvest was significantly decreased, and the extent of reduction was positively related to the anti-aging ability. Consequently, the seed cotton yield of PSAG12-IPT lines (OE-37 and OE-38) with strong or moderate anti-aging ability decreased due to less opened bolls, while the line (OE-30) with weak anti-aging ability had a comparable yield to that of wild type. The fiber length uniformity of PSAG12-IPT cotton lines was significantly higher than that of wild type. Compared with wild type, the micronaire results of transgenic lines revealed an increasing trend, and fiber length, breaking strength, and breaking elongation was not significantly different from that of wild type.

Key words: cotton, PSAG12-IPT, cytokinin, yield, fiber quality

图1

质粒PSG529结构图"

图2

转PSAG12-IPT基因棉花植株的PCR检测 A: 叶片中IPT基因的PCR结果; B: 叶片中NPT II基因的PCR结果; C: 根系中NPT II基因的PCR结果。"

图3

棉花叶片衰老阶段PSAG12-IPT基因在转基因株系中的表达分析 柱上不同小写字母表示在0.05水平差异显著。"

图4

转PSAG12-IPT基因对棉花叶片衰老阶段细胞分裂素含量的影响 柱上不同小写字母表示在0.05水平差异显著。"

表1

不同转基因棉花株系叶片细胞分裂素的聚类分析"

类群
Cluster		 株系代号
Code of line		 株系个数
Number of
lines		 频率
Frequency
(%)		 类平均标准差
Mean of cluster
(nmol g-1)		 变幅
Range
(nmol g-1)
I OE-9, OE-30 2 22.22 75.66 c 75.45-75.88
II OE-17, OE-19B, OE-19L, OE-22, OE-27, OE-37 6 66.67 87.43 b 82.23-93.19
III OE-38 1 11.11 113.62 a 113.62

图5

PSAG12-IPT基因在转基因棉株叶片中相对表达量变化动态"

图6

转基因株系和野生型棉株叶片细胞分裂素含量变化动态"

图7

转基因株系和野生型棉株叶片叶绿素含量变化动态"

图8

转基因株系和野生型棉株叶片可溶性蛋白含量变化动态"

表2

PSAG12-IPT基因导入对棉花农艺性状的影响"

株系
Line		 2011 2012
株高
Plant height
(cm)		 果枝始节
First node
producing fruiting branch		 果枝数
Number of fruiting branch		 株高
Plant height
(cm)		 果枝始节
First node
producing fruiting branch		 果枝数
Number of fruiting branch
冀合321 Jihe 321 103.3 a 6.3 b 14.0 a 134.5 a 7.6 b 16.9 a
OE-30 91.7 bc 8.0 a 12.3 a 127.1 a 9.5 a 16.5 a
OE-37 88.0 c 8.0 a 12.0 a 129.8 a 8.3 a 16.2 a
OE-38 100.3 b 8.3 a 13.0 a 123.4 a 9.6 a 15.0 a

表3

PSAG12-IPT基因导入对棉花产量和产量构成因素的影响"

年份
Year		 株系
Line		 单株成铃数
Boll number per plant		 铃重
Boll weight
(g)		 衣分
Lint percentage
(%)		 吐絮率
Boll opening percentage
(%)		 籽棉产量
Seed cotton yield
(kg hm-2)
2011 冀合321 Jihe 321 12.4 b 4.2 ab 37 a 94.4 a 1257.0 a
OE-30 18.6 a 4.6 a 36 a 71.5 b 1364.8 a
OE-37 12.3 b 4.0 b 40 a 53.2 bc 814.9 b
OE-38 17.7 a 3.8 b 34 a 37.9 c 401.3 c
2012 冀合321 Jihe 321 24.2 a 4.3 a 35 a 49.9 a 1554.9 a
OE-30 24.0 a 4.2 a 33 a 30.0 b 1605.9 a
OE-37 21.0 a 4.0 a 33 a 22.7 c 1405.1 b
OE-38 23.6 a 3.7 a 32 a 8.6 d 1335.5 b

表4

PSAG12-IPT基因导入对棉花品质的影响"

株系
Line		 纤维长度
Fiber length
(mm)		 整齐度指数
Uniformity index
(%)		 马克隆值
Micronaire value		 断裂伸长率
Breaking elongation
(%)		 断裂比强度
Breaking strength
(cN tex-1)
冀合321 Jihe 321 29.2 a 82.9 c 4.1 b 6.5 a 29.5 a
OE-30 31.6 a 84.7 b 4.4 ab 6.2 a 30.6 a
OE-37 30.5 a 84.0 b 4.3 b 6.4 a 30.5 a
OE-38 29.5 a 85.3 a 4.7 a 6.3 a 29.8 a
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