中国农业科学 ›› 2022, Vol. 55 ›› Issue (8): 1503-1517.doi: 10.3864/j.issn.0578-1752.2022.08.003
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()
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:
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()
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。
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. 陆地棉遗传标准系TM-1的特性及其耐冷性[J]. 中国农业科学, 2022, 55(8): 1503-1517.
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. Characteristics and Cold Tolerance of Upland Cotton Genetic Standard Line TM-1[J]. Scientia Agricultura Sinica, 2022, 55(8): 1503-1517.
表1
供试材料来源及系谱"
名称 Name | 来源或培育单位 Origin or the breeding group | 亲本组合 Parental combinations |
---|---|---|
TM-1 | 国家棉花种质中期库(安阳) National Medium-term Genebank of Cotton Germplasm Resource in China (Anyang) | 品系D&PL-14 The line D&PL-14 |
豫2067 Yu 2067 | 河南省农业科学院经济作物研究所 Institute of Economic Crops, Henan Academy of Agricultural Sciences | 中棉所12/豫植177 CRI12/Yuzhi 177 |
衡棉3号 Hengmian 3 | 河北省农林科学院旱作农业研究所 Dry Farming Agricultural Research Institute, Hebei Academy of Agricultural and Forestry Sciences | 衡9273/GK12 Heng 9273/GK12 |
中棉所41 CRI41 | 中国农业科学院棉花研究所 Institute of Cotton Research of Chinese Academy of Agricultural Sciences | 中棉所23导入国产Bt和CpTI双价抗虫基因 Introduction of Bt and CpTI genes into CRI23 |
中棉所35 CRI35 | 中国农业科学院棉花研究所 Institute of Cotton Research of Chinese Academy of Agricultural Sciences | 中23021/(中棉所12×川1704) Zhong 23021/(CRI12×Chuang 1704) |
表2
11个耐冷相关基因在转录组中的表达倍数和实时灾光定量引物"
基因编号 Gene ID | 基因 Gene | 基因注释 Gene functional annotations | 表达倍数 log2Ratio (Treatment/Control) | 正向引物 Forward primer sequence (5′-3′) | 反向引物 Reverse primer sequence (5′-3′) |
---|---|---|---|---|---|
CotAD_50094 | ZAT11 | 锌指蛋白ZAT11 Zinc finger protein ZAT11 | 2.714 | AACGGAGAACCAACCACCAG | CCCAAGGCTTGTCCGATTGA |
CotAD_58358 | COR47 | 脱水素COR47 Dehydrin COR47 | 2.129 | GTTAGCGGTGAAGGAGCAGT | ACTCGGTTACGATCACCTCC |
CotAD_22633 | ARG2 | 吲哚-3-乙酸诱导蛋白ARG2 Indole-3-acetic acid-induced protein ARG2 | 6.762 | GCTACGGTGGCCAAGAAAAC | TTGGGACAGTTCTCGGGTCT |
CotAD_74365 | PBP1 | 钙信号转导的钙结合蛋白PBP1 Calcium-binding protein PBP1 | 5.134 | CTTGAAGAGGAACGCTGCTG | TTCCATTTCACTGAGAGCCCC |
CotAD_45286 | CIPK6 | CBL相互作用蛋白激酶 CBL-interacting serine/threonine-protein kinase 6 | 4.209 | GTGTACGACTGCAGGGTCAA | ACCAGAAACGACGGTGTCAA |
CotAD_09571 | MPK3 | 丝裂原活化蛋白激酶3 Mitogen-activated protein kinase 3 | 2.244 | CTGCTTGGCACTCCAACTGA | TGTTGACGAGGGTATGCAGG |
CotAD_12521 | EXPA8 | 细胞壁伸展蛋白A8 Expansin-A8 | -7.391 | AGGGTTTGGGACCAACACAG | AGCAGGCAAGCACCATTTTG |
CotAD_76943 | TRG-31 | 可能的水通道蛋白PIP 7a型 Probable aquaporin PIP-type 7a | 3.050 | AGGTATGGTGCTTTGGGTGG | CTTGGCATCAGTGGCTGAGA |
CotAD_17081 | WRKY46 | 可能的WRKY转录因子46 Probable WRKY transcription factor 46 | 2.684 | CCTCGCTTAGCACGATGGAA | GCCAGAGCGAACCCTAAGTT |
CotAD_55002 | NAC100 | 含NAC结构域的蛋白100 NAC domain-containing protein 100 | 4.545 | ACTTCAGTGCCGCTGCTATT | TCCCCCATCTTGGCTTTGTC |
CotAD_56948 | poxN1 | 过氧化物酶N1 Peroxidase N1 | 3.234 | TGGAGGACACACCATAGGGA | TTGAGGCACGAATGCTGAGT |
表3
供试材料的农艺性状"
名称 Name | 株高 Plant height (cm) | 单株果枝数 Number of fruit branches per plant | 单株铃数 Number of bolls per plant | 铃重 Single boll weight (g) | 衣分 Lint percentage (%) | 籽棉产量 Unginned cotton yield (kg·hm-2) |
---|---|---|---|---|---|---|
TM-1 | 94.60Aa | 12.80A | 10.40A | 6.55Aa | 39.18Aa | 2791.50A |
中棉所35 CRI35 | 84.40Ab | 9.10B | 7.80B | 5.65Ab | 38.66Aa | 2347.20B |
表5
TM-1子叶期的耐冷性鉴定"
品种 Variety | 处理 Treatments | 相对叶绿素含量 The relative chlorophyll content (SPAD value) | 耐冷指数 Index of cold tolerance (%) | 耐冷级别 Level of cold tolerance |
---|---|---|---|---|
TM-1 | CK | 53.22Ab | ||
4℃ 4 d | 52.33Ab | |||
CK恢复7 d Recovery for 7 d of CK | 60.2Aa | |||
4℃4 d恢复7 d Recovery for 7 d after 4℃4 d | 53.91Ab | 85.32Aa | 抗冷Chilling resistant | |
豫2067 Yu 2067 | CK | 52.61Ab | ||
4℃4 d | 51.76Ab | |||
CK恢复7 d Recovery for 7 d of CK | 60.8Aa | |||
4℃4 d恢复7 d Recovery for 7 d after 4℃4 d | 54.16Ab | 76.55Ab | 抗冷Chilling resistant | |
衡棉3号 Hengmian 3 | CK | 51.12Ab | ||
4℃4 d | 41.14B | |||
CK 恢复7 d Recovery for 7 d of CK | 60.6Aa | |||
4℃4 d恢复7 d Recovery for 7 d after 4℃4 d | 41.48B | 21.62B | 冷敏感Chilling sensitive |
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