中国农业科学 ›› 2022, Vol. 55 ›› Issue (8): 1503-1517.doi: 10.3864/j.issn.0578-1752.2022.08.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

陆地棉遗传标准系TM-1的特性及其耐冷性

王俊娟(),陆许可,王延琴,王帅,阴祖军,付小琼,王德龙,陈修贵,郭丽雪,陈超,赵兰杰,韩迎春,孙亮庆,韩明格,张悦新,范亚朋,叶武威()   

  1. 中国农业科学院棉花研究所/棉花生物学国家重点实验室/农业农村部棉花遗传改良重点开放实验室,河南安阳 455000
  • 收稿日期:2021-12-16 接受日期:2022-02-11 出版日期:2022-04-16 发布日期:2022-05-11
  • 联系方式: 王俊娟,Tel:15093920872;E-mail: wjj2004liyuan@sina.com。
  • 基金资助:
    财政部和农业农村部:国家现代农业产业技术体系资助(CARS-15-02);中央级公益性科研院所基本科研业务费专项(1610162021036);转基因重大专项(2016ZX08005004-005)

Characteristics and Cold Tolerance of Upland Cotton Genetic Standard Line TM-1

WANG JunJuan(),LU XuKe,WANG YanQin,WANG Shuai,YIN ZuJun,FU XiaoQiong,WANG DeLong,CHEN XiuGui,GUO LiXue,CHEN Chao,ZHAO LanJie,HAN YingChun,SUN LiangQing,HAN MingGe,ZHANG YueXin,FAN YaPeng,YE WuWei()   

  1. Institute of Cotton, Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology/Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture and Rural Affairs, Anyang 455000, Henan
  • Received:2021-12-16 Accepted:2022-02-11 Published:2022-04-16 Online:2022-05-11

摘要:

【目的】通过系统调查TM-1农艺性状及纤维品质,研究芽期和苗期的耐冷性,以及对其在低温胁迫条件下的耐冷相关基因进行实时荧光定量分析,深入挖掘其耐冷机理,为该种质利用提供理论依据。【方法】在田间,以中棉所35为对照,以人工调查方式对TM-1进行表型鉴定,依据国际校准棉花标准(HVICC)测定纤维品质,同时采用卡那霉素筛选和分子鉴定进行Bt检测。以抗冷品种豫2067和冷敏感品种衡棉3号为对照,对TM-1芽期和子叶期进行耐冷性鉴定,4℃低温处理,然后正常条件下恢复生长7 d,调查芽期相对子叶平展率及子叶期植株受胁迫后的冷害级别,计算冷害指数和耐冷指数;使用便携式叶绿素仪进行叶片活体测定,以SPAD值代表相对叶绿素含量;在三叶期进行4℃低温处理24 h,利用实时荧光定量方法进行耐冷相关基因在叶片中的表达测定。【结果】表型鉴定结果显示,TM-1叶片较大,颜色深绿,生育期较长,约135 d,霜前籽棉产量为2 791.50 kg·hm-2,株高94.60 cm,霜前籽棉产量、株高、单株果枝数和单株结铃数均高于中棉所35,其他农艺性状与中棉所35接近,纤维品质中等水平。卡那霉素和试纸检测结果表明TM-1与中棉所35均不含Bt。芽期耐冷性鉴定结果表明,与对照处理相比,TM-1叶片的相对叶绿素含量极显著下降,株高极显著降低,低温胁迫极显著抑制了棉花下胚轴的伸长,并且抑制了叶片叶绿素的合成;低温处理后,TM-1的主根受到了伤害,恢复生长后侧根却比较发达,侧根生长优于对照。TM-1芽期耐冷级别达到高抗冷。苗期抗冷性鉴定结果表明,与对照相比,TM-1的相对叶绿素含量显著下降,株高极显著降低。TM-1子叶期耐冷指数为85.32%,显著高于抗冷材料豫2067,耐冷级别达到抗冷。TM-1三叶期受低温胁迫24 h后,在叶片中,有9个耐冷相关基因上调表达,上调表达倍数显著或极显著高于衡棉3号;其中,脱水素基因在叶片中上调表达,表达倍数与其在豫2067叶片中的表达倍数相近,是衡棉3号叶片中表达倍数的4.69倍,LEA3在TM-1叶片中的上调表达倍数显著高于豫2067和衡棉3号。总之,TM-1属于中晚熟品系,纤维品质中等,不含Bt;芽期和子叶期均比较抗冷,属于比较抗冷的类型;与耐冷相关的基因在TM-1的叶片中受低温诱导表达,拥有特别的表达模式。【结论】TM-1具有稳定的农艺性状,纤维品质达中等水平,不含Bt,可用作转外源基因的良好受体;因为其良好的耐冷特性,TM-1可以作为棉花耐冷性育种改良的重要亲本来源和基因克隆的基因源。

关键词: 棉花, 低温胁迫, TM-1, 耐冷性

Abstract:

【Objective】We systematically investigated the major agronomic traits and cold tolerance of accession TM-1 at the bud and seedling stages. The relative expressions of cold tolerance-related genes were analyzed by the qRT-PCR method. The cold tolerance mechanism of TM-1 was further discussed, which provides the theoretical basis for the breeding utilization of TM-1.【Method】The major agronomic traits of TM-1 were manually investigated in the field using variety CRI35 as the control. The fiber quality was assessed by an international calibrated cotton standard (HVICC), and the insect resistance (Bt) was detected by kanamycin screening and molecular detection technologies. For the cold tolerance testing, two contrasting accessions, cold-resistant accession Yu 2067 and cold-sensitive variety Hengmian 3 were set as controls, respectively. The cold resistance of TM-1 at bud stage and cotyledon stage was identified, treated at 4℃ and then recovered under normal conditions for 7 days, and the relative cotyledon spreading rate and the cold injury levels of plants were investigated, and cold injury indexes and cold resistance indexes were calculated. The portable chlorophyll meter was used for in vivo testing the leaf relative chlorophyll content (represented by SPAD value). The expressions of cold tolerance-related genes in leaves were measured by qRT-PCR method. 【Result】 The leaves of TM-1 were large and dark green. The pre-frost seed cotton yield was 2 791.50 kg·hm-2, and the plant height was 94.60 cm. The growth period was about 135 days, and the yield, plant height, fruit branch number per plant, boll number per plant were higher than CRI35, while other agronomic traits were similar to CRI35. TM-1 had medium fiber quality. The test results of kanamycin and test paper showed that TM-1 did not contain the Bt like CRI35. Identification results of cold tolerance at bud stage showed that compared with the control treatment, the relative chlorophyll content and plant height of TM-1 decreased significantly. Low-temperature stress significantly inhibited hypocotyl elongation and chlorophyll synthesis in cotton leaves. Under low-temperature treatment, the taproots of TM-1 were damaged, but the lateral roots were more developed than those of the control. The cold tolerance level of TM-1 reached high cold resistance at the bud stage. Identification of cold tolerance at the cotyledon stage showed that the relative chlorophyll content and plant height of TM-1 decreased significantly compared with the control. The cold tolerance index of TM-1 at the cotyledon stage was 85.32%, which was significantly higher than Yu 2067, and the tolerance level of TM-1 reached cold resistance at the cotyledon stage. After the treatment of low-temperature stress for 24 h at the trefoil stage, nine genes were up-regulated in the TM-1 leaves, and their up-regulated expression folds were significantly higher than those of cold-sensitive accession. Dehydrin gene was up-regulated in TM-1 leaves, and the expression fold was similar to that in the leaves of Yu 2067, which was 4.69 times that in the leaves of Hengmian 3. The expression fold of the LEA3 gene in TM-1 leaves was significantly higher than that of Yu 2067 and Hengmian 3. 【Conclusion】 Accession TM-1 has stable agronomic characters and the medium fiber quality. It can be used as an ideal receptor for transferring exotic genes because without Bt. TM-1 can also be used as an important parent for cotton breeding and a gene source for cloning genes because of its good cold tolerance.

Key words: cotton, low temperature stress, TM-1, cold tolerance