作物杂志, 2026, 42(3): 126-131 doi: 10.16035/j.issn.1001-7283.2026.03.017

遗传育种·种质资源·生物技术

烟草两种质源胞质雄性不育系营养元素差异分析

许杰,1, 郑昀晔1, 陈学军2, 王国平1, 耿世兵1, 牛永志1, 张立猛,1,2

1 玉溪中烟种子有限责任公司653100云南玉溪

2 云南省烟草农业科学研究院650021云南昆明

Difference Analysis of Nutrient Elements in Two Cytoplasmic Male Sterile Lines of Tobacco

Xu Jie,1, Zheng Yunye1, Chen Xuejun2, Wang Guoping1, Geng Shibing1, Niu Yongzhi1, Zhang Limeng,1,2

1 Yuxi Zhongyan Tobacco Seed Company, Yuxi 653100, Yunnan, China

2 Yunnan Academy of Tobacco Agricultural Sciences, Kunming 650021, Yunnan, China

通讯作者: 张立猛,主要从事烟草绿色防控和种子技术研究,E-mail:zlm.d@163.com

收稿日期: 2025-02-11   修回日期: 2025-06-6   网络出版日期: 2025-07-09

基金资助: 云南省烟草公司重大项目“烟草品种‘育繁推’一体化平台构建研究及应用”(2022530000241016)
“烟草花器官发育关键调控机理解析和技术应用”(2025530000241002)

Received: 2025-02-11   Revised: 2025-06-6   Online: 2025-07-09

作者简介 About authors

许杰,主要从事烟草种子技术研究,E-mail:xujie2025@126.com

摘要

为比较不同质源烟草雄性不育系及其保持系的产量和营养元素含量差异,以不同胞质来源的雄性不育系Nta(sua)S K326 [(sua)S]、Nta(gla)S K326 [(gla)S]和保持系K326(MF K326)为材料,研究了3个基因型雌雄蕊形态、种子产量以及不同时期和部位营养元素含量。结果表明,相较于MF K326正常雌雄蕊,(gla)S的雌蕊正常,雄蕊退化,花丝短小,花药皱缩;(sua)S雌蕊正常或变扁、开裂,雄蕊雌化,花丝顶端长出柱头状结构。MF K326和(gla)S的种子产量分别是(sua)S产量的1.51和1.38倍。不同基因型总氮、镁、硼和铜含量在不同时期的叶片和花/果中无显著差异;MF K326钾含量在3个时期的叶和雌雄蕊中显著高于2个不育系,除青果期叶片外2个不育系磷酸盐含量在不同时期始终低于MF K326,铁和钙在不育系不同时期的花/果中积累;(sua)S锌含量在盛花期和青果期叶片中低于另外2个基因型,锰含量在不同发育时期和雌雄蕊中均最低,而(gla)S在不同发育时期锰含量均最高。本研究初步分析了营养元素与种子产量的关系,为可能的(sua)S繁种生产及产量改良提供了参考。

关键词: 烟草; 不育胞质; 营养元素

Abstract

In order to compare the differences in yield and nutrient element contents between different cytoplasmic male sterile (CMS) lines and their maintainer lines of tobacco, we used CMS lines with different cytoplasmic sources, Nta(sua)S K326 [(sua)S], Nta(gla)S K326 [(gla)S], and the maintainer line K326 (MF K326), as materials. We investigated the morphology of stamens and pistils, seed yield, and the contents of nutrient elements in different parts and at different stages of these three genotypes. The results showed that compared with the normal stamens and pistils of MF K326, the pistils of (gla)S were normal, while the stamens were degenerated, with short filaments and shrunken anthers. The pistils of (sua)S were normal or flattened or split, and the stamens were feminized, with stigma-like structures growing at the tips of the filaments. The seed yields of MF K326 and (gla)S were 1.51 and 1.38 times that of (sua)S, respectively. There were no significant differences in the total nitrogen, magnesium, boron, and copper contents among different genotypes in leaves and flowers or fruits at different stages. The potassium content in MF K326 was significantly higher than that in the two CMS lines in leaves and stamens and pistils at three stages. The phosphate content of the two CMS lines was always lower than that of MF K326 at different stages, except for the leaves at the green fruit stage. Iron and calcium accumulated in the flowers/fruits of the CMS lines at different stages. The zinc content of (sua)S was lower than that of the other two genotypes in leaves at the full bloom and green fruit stages. The manganese content of (sua)S was the lowest in flowers/fruits at different developmental stages and in stamens and pistils, while the manganese content of (gla)S was the highest at different developmental stages. This study preliminarily analyzed the relationship between nutrient elements and seed yield, providing a reference for potential seed production and yield improvement of (sua)S.

Keywords: Tobacco; Sterile cytoplasm; Nutrient elements

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本文引用格式

许杰, 郑昀晔, 陈学军, 王国平, 耿世兵, 牛永志, 张立猛. 烟草两种质源胞质雄性不育系营养元素差异分析. 作物杂志, 2026, 42(3): 126-131 doi:10.16035/j.issn.1001-7283.2026.03.017

Xu Jie, Zheng Yunye, Chen Xuejun, Wang Guoping, Geng Shibing, Niu Yongzhi, Zhang Limeng. Difference Analysis of Nutrient Elements in Two Cytoplasmic Male Sterile Lines of Tobacco. Crops, 2026, 42(3): 126-131 doi:10.16035/j.issn.1001-7283.2026.03.017

烟草是中国重要的经济作物之一,为保证品种纯度,充分发挥杂种优势,烟草胞质雄性不育系得到广泛应用,雄性不育杂交种种植已超过全国烤烟总种植面积的50%[1]。目前,烟叶生产上主栽品种(MS K326、MS云烟87)的不育胞质均来自于香甜烟草(Nicotiana suaveolens)[以(sua)S表示,下同][2]。该类型的雄性不育系雌蕊畸形率极高(>50%),授粉后落花落果严重,对种子生产造成极大损失。前人[3]研究表明,(sua)S导致种子产量降低,影响了烟草种子繁育效率。且长期使用单一不育胞质会导致遗传基础脆弱,使烟叶生产存在潜在风险[4]

N.glauca可以作为替代N.suaveolens的潜在来源[5]。近年来,国内研究人员获得了源自N.glauca [以(gla)S表示,下同]的不育胞质[2]。不同来源的烟草不育胞质具有不同的败育特点和胞质效应[6]。目前,关于不同不育胞质不育机理的研究主要集中在表型、细胞学、生理生化和以线粒体orf为主的分子机理等方面,对矿质营养研究比较少。

本文以(gla)S与(sua)S 2种来源的不育系与其保持系MF K326为材料,研究了雌雄蕊形态、种子产量和不同时期烟草叶/花/果和雌、雄蕊中大、中、微量元素含量差异,初步探讨了营养元素与不育系不育形成及产量的关系,为进一步促进不育胞质挖掘和筛选以及可能的(sua)S繁种生产和产量提高提供理论依据。

1 材料与方法

1.1 试验材料

Nta(sua)S K326胞质[以(sua)S表示,下同]、Nta(gla)S K326[以(gla)S表示,下同]和保持系K326(以MF K326表示,下同)的种子由云南省烟草农业科学研究院育种室提供。

1.2 试验方法

试验于2023年9月至2024年3月在玉溪中烟种子有限责任公司西双版纳冬繁基地开展。2023年9月初育苗,11月初移栽,田间常规管理。施肥方法:有机肥(油枯型,总养分≥8%)1500 kg/hm2(100 kg/亩),整地时均匀撒施于土壤;复合肥(N:P2O5:K2O=12:8:24)990 kg/hm2(66 kg/亩),整地时撒施630 kg/hm2(42 kg/亩)作为基肥,剩余的分别于移栽、团棵、旺长和现蕾期时按90 kg/hm2(6 kg/亩)随水施入烟株根际;钙镁磷(0-18-0)按375 kg/hm2(25 kg/亩)于团棵期撒施于烟株根际;硫酸锌(≥21.5%)15 kg/hm2(1 kg/亩),按7.5 kg/hm2(0.5 kg/亩)分别于团棵期和旺长期随水施入烟株根际;硼砂30 kg/hm2(2 kg/亩)(≥95%),分别于移栽、团棵、旺长和现蕾期按7.5 kg/hm2(0.5 kg/亩)随水施入烟株根际。

分别于初花期、盛花期和青果期采集MF K326、(gla)S、(sua)S烟叶和花/果,在105 ℃杀青15 min,并在60 ℃烘干6 h,粉碎后过60目筛备用。于盛花期采集MF K326、(gla)S、(sua)S S1时期(小孢子,裂口分化)花蕾和S8时期(花始开期,花粉成熟)雌雄蕊混合样(50朵花剥出全部雌雄蕊混合在一起),按上述方法杀青粉碎后过筛备用。

于盛花期开始人工授粉,共计授粉5次,每株授粉150~250朵花。分批次成熟采收,晒干后脱粒,通过筛选和风选去除杂质,过45目筛后称重,每个处理100株,同期种植,分3次重复。按照14 250株/hm2(950株/亩)换算成单位面积产量。

1.3 测定项目与方法

按照《烟草及烟草制品 总氮的测定 连续流动法》(YC/T 161-2002)[7]测定总氮含量;按照《烟草及烟草制品 钾的测定 连续流动法》(YC/T 217-2007)[8]检测钾含量;按照《烟草及烟草制品 钙的测定 原子吸收法》(YC/T 174-2003)[9]检测钙含量;按照《烟草及烟草制品 镁的测定 原子吸收法》(YC/T 175-2003)[10]检测镁含量;按照微波消解-ICP方法测定磷酸盐含量;按照微波消解-ICP MS方法测定硼含量;按照微波消解-ICP-MS方法测定铜、锌和锰含量;按照《森林植物与森林枯枝落叶层全硅、铁、铝、钙、镁、钾、钠、磷、硫、锰、铜、锌的测定》(LY/T 1270-1999)[11]测定铁含量。

1.4 数据处理

采用DPS 7.05对数据进行分析,采用Excel 2010进行绘图。

2 结果与分析

2.1 不同质源胞质不育系及保持系的雌雄蕊形态差异

通过比较3个基因型的雌雄蕊(图1)可以看出,(gla)S的雌蕊形态与MF K326没有差异,但是雄蕊退化,花丝短小,花药皱缩;而(sua)S的雌雄蕊形态则与MF K326差异巨大,柱头、子房和花柱均畸形,雄蕊表现出心皮化,花丝顶端长出了柱头状结构(如图1箭头所示),雌蕊中大约70%~80%表现出如图1所示的结构。

图1

图1   不同基因型雌雄蕊形态差异

Fig.1   Morphological differences in pistils and stamens of different genotypes


2.2 不同质源胞质不育系及保持系的种子产量差异

对不同质源胞质不育系进行人工授粉,保持系自交结实,种子成熟后进行采收并统计3个不同基因型的种子产量,结果(图2)显示,保持系MF K326产量最高,不育系(gla)S居中,不育系(sua)S最低,MF K326和(gla)S的种子产量分别是(sua)S的1.51和1.38倍,且差异达到极显著水平。经田间观察,可能原因是(sua)S胞质导致雌蕊畸形后,部分子房无法完成受精,导致落花落果或蒴果不饱满,从而造成种子产量降低。

图2

图2   不同基因型产量比较

不同大写字母代表差异极显著(P < 0.01)。

Fig.2   Comparison of yields among different genotypes

Different capital letters indicate extremely significant differences (P < 0.01).


2.3 不同质源胞质不育系及保持系不同发育时期营养元素含量差异

营养元素作为酶和电子载体的重要组分,在植物生理生化过程中起着重要作用[12]。3个不同基因型烟草在不同时期营养元素差异见表1。由表1可知,总氮、镁、硼和铜含量在不同时期的叶片和花/果中基因型间无显著差异。MF K326磷酸盐含量在初花期和盛花期的花、叶和青果期的果中均显著高于另外2个基因型,在青果期的叶中低于另外2个基因型;MF K326钾含量在3个时期的叶中要明显高于另外2个基因型,在花/果中无显著差异;钙含量在3个时期的叶中MF K326要明显高于另外2个基因型,但是在3个时期的花/果中,MF K326的钙含量要明显低于另外2个基因型;(sua)S的铁含量在不同时期的叶中最低,在花/果中含量显著高于MF K326,而与(gla)S差异不显著;(sua)S的锌含量在盛花和青果期叶中均显著低于另外2个基因型,在其他时期的叶/花/果中3个基因型差异均不显著;(sua)S的锰含量在不同时期的叶和花/果中均最低,而(gla)S的锰含量在不同时期的叶和花/果中含量均最高,差异显著。

表1   不同基因型不同生育期营养元素差异

Table 1  Differences in nutrient elements of different genotypes at various growth stages

生育期
Growth stage
器官
Organ
基因型
Genotype
总氮
N
(%)
磷酸盐
Phosphate
(%)

K
(%)

Ca
(%)

Mg
(%)

B
(mg/kg)

Fe
(g/kg)

Cu
(mg/kg)

Zn
(mg/kg)

Mn
(mg/kg)
初花期
Early flowering stage
MF K3263.01a0.78a5.34a2.48a0.30a24.69a0.54a9.16a33.75a29.00a
(gla)S2.79a0.67b4.27b1.90b0.31a22.83a0.45b9.12a29.99a31.97a
(sua)S2.87a0.64b4.88ab1.94ab0.31a22.62a0.37c7.83a28.78a16.32b
MF K3264.52a1.59a4.21a0.46b0.39a26.81a0.15b24.75a55.43a32.24b
(gla)S4.19a1.46b4.08a0.63a0.41a25.65a0.29a23.49a50.68a44.41a
(sua)S4.31a1.40b4.03a0.59a0.41a25.92a0.26a24.10a53.26a27.30b
盛花期
Full flowering stage
MF K3261.77a0.65a4.22a2.38a0.24a24.25a0.71a8.32a28.41a33.31b
(gla)S1.58a0.53b3.24b1.90b0.28a22.84a0.68a9.67a25.74a39.46a
(sua)S1.82a0.52b3.43b1.99b0.29a22.59a0.49b8.80a17.57b21.86c
MF K3262.96a0.85a3.46a0.58b0.25a34.00a0.25b16.29a38.06a32.23b
(gla)S3.20a0.70c3.44a0.68a0.30a32.33a0.33a17.31a42.55a42.10a
(sua)S3.15a0.79b3.55a0.63ab0.29a32.86a0.30a16.93a41.15a26.09b
青果期
Green fruits stage
MF K3261.26a0.80b3.74a3.10a0.34a27.02a1.09a12.37a35.30a50.09b
(gla)S1.27a0.84ab3.02b2.61b0.40a28.53a0.83b12.55a38.79a62.61a
(sua)S1.43a0.89a2.89b2.87ab0.40a26.58a0.52c11.97a25.41b29.31c
MF K3262.71a1.00a3.26a0.35b0.24a35.58a0.28b21.61a47.60a32.72ab
(gla)S2.64a0.92b3.22a0.38b0.25a34.01a0.38a22.35a47.77a36.55a
(sua)S2.90a0.89b3.31a0.47a0.26a36.10a0.36a20.55a44.98a28.17b

不同小写字母表示处理间差异显著(P < 0.05)。下同。

Different lowercase letters indicate significant differences among treatments (P < 0.05). The same below.

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2.4 不同质源胞质不育系及保持系雌雄蕊营养元素含量差异

前人[13]研究发现,烟草雄性不育可能与营养代谢紊乱而间接抑制雄性器官发育有关。本研究进一步检测了盛花期(S1和S8)雌雄蕊混合样中磷酸盐、铁、钾、钙、锌及锰含量。结果(表2)表明,MF K326在2个时期的磷酸盐和钾含量均显著高于另外2个基因型;MF K326在2个时期的钙和铁含量均低于另外2个基因型;(gla)S在2个时期的锰含量均高于另外2个基因型;3个基因型在不同时期的锌含量差异不显著。说明(sua)S基因型导致雌雄蕊中钙和铁含量升高,磷酸盐、钾和锰含量降低。

表2   不同基因型烟草不同花期雌雄蕊主要营养元素差异

Table 2  Differences in major nutrient elements in pistils and stamens of tobacco with different genotypes during flowering stage

基因型Genotype磷酸盐Phosphate (%)钾K (%)钙Ca (%)铁Fe (g/kg)锌Zn (mg/kg)锰Mn (mg/kg)
MF K326-S11.47a3.35a0.29b0.10c53.10a29.80b
(gla)S-S11.43b3.15c0.33a0.14a52.13a32.43a
(sua)S-S11.41b3.25b0.32a0.12b53.57a30.47b
MF K326-S81.78a3.13a0.25b0.10b48.43a26.73b
(gla)S-S81.53b3.03b0.31a0.11b49.13a29.30a
(sua)S-S81.59b2.95c0.31a0.14a48.30a24.00c

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3 讨论

有研究[3]表明,尽管细胞质遗传因子只占烟草遗传变异的很小一部分,但用其他Nicotiana物种的细胞质替代原有细胞质会产生超出修改雄性生殖器官本身的影响。相较于正常雄性可育品种,外源细胞质对细胞质雄性不育的农业性状的影响,如果存在,通常并非总是有害的,但其变化范围从几乎不可察觉到相当大[14]。Berbeć等[15]研究了14个不同来源不育胞质和它们的雄性可育品种‘Wiślica’的农艺性状和经济性状,结果显示,cms raimondiicms megalosiphoncms occidentalis的叶片尺寸减小,cms plumbaginifoliacms tabacum (glutinosa)cms eastii的植物明显更矮,cms knightianacms raimondiicms eastiicms tabacum (glutinosa)cms plumbaginifolia的开花时间延迟。早在1955年,Izard等[16]就发现N.suaveolens胞质来源的雄性不育系花冠正常,雌蕊正常或缩短,具多裂柱头;雄蕊缺失,呈柱头状或心皮状;花的变异程度因核基因型和环境而异。Berbeć[17]在1966年发现,N.glauca正常花冠,雄蕊缺失或未发育完全。在本研究中,2个不同质源胞质不育系及其保持系的雌雄蕊性状与前人[15-17]研究结果一致,说明该现象是由于细胞质来源不同导致的。

由于来源于N.suaveolens的不育胞质与其保持系在烟株高度、开花天数、叶片数量、叶面积以及单叶重、产量、产值等方面没有任何劣势[15],普遍认为在农业上适宜,并且在全世界的杂交育种和种子生产中使用最广泛[18]。但是,由于N.suaveolens不育胞质与特定核基因型组合时降低了雌蕊育性,导致杂交种子产量严重下降,且N.suaveolens不育胞质增加了对某些疾病如马铃薯Y病毒病和蛙眼病的敏感性,因此,育种家一直在积极寻找替代cms suaveolens的不育胞质[3]。Amankwa等[5]的研究表明,N.glauca有可能成为替代cms suaveolens的潜在不育胞质来源,N.glauca不育胞质对植物生长、产量和化学成分没有负面影响,与cms suaveolens相比,杂交种子的产量有所提高,与保持系相比,叶片产量略有下降,还原糖、绿原酸和芸苔素含量增加,开花天数减少。本研究结果表明,(sua)S显著降低了MF K326种子产量,这与N.suaveolens不育胞质影响了雌蕊的育性有关,(sua)S柱头、花柱和子房均畸形可能是种子产量降低的直接原因;而(gla)S雌蕊正常,种子产量显著高于(sua)S。有研究[19]表明,采用了N.glauca cms系统的不育系或杂交种已经实现大规模的商业种植。

早在1987-1989年,于绍夫等[20]检测了果树10个树种、82个品种花粉中的15种矿质元素,结果显示,钾含量最高,磷、镁、钙次之,其后是铁、锰、硼、铜、锌。王开发等[21]在1998年测定了烟草花粉中主要矿质元素,结果显示,磷(2 mg/g)>镁(0.32 mg/g)>钙(0.16 mg/g)>铁(0.1 mg/g)>锰(0.02 mg/g)。因此,MF K326不同时期叶中钾、磷酸盐和钙含量高于2个不育系,可能是花粉发育过程中需要较多K、P和Ca所致。花粉发育是一个高耗能的过程,多种作物花粉败育的发生伴随着三磷酸腺苷(ATP)含量的降低。在玉米、油菜、大豆和烟草等物种中,不育系ATP含量显著低于可育系[22]。花粉粒内壁畸形和ATP酶活性差异可能是导致K型小麦花粉粒败育的重要原因[23]。洋葱T型胞质不育系花粉成熟早期ATP含量仅为保持系的1%,在其他时期含量均最低[24]。在本研究中,MF K326磷酸盐含量不同时期(包括花朵2个时期,除青果期叶)始终高于2个不育系,可能与不育系ATP含量降低有关。细胞质雄性不育(CMS)通过线粒体功能障碍阻止功能性花粉的产生,雄蕊雌蕊化受核质互作的调控,这种核质互作通过一种细胞内信号通路——线粒体逆向信号(MRS)发生[25],而铁是线粒体内电子传递链上许多酶的辅助因子[26]。在本研究中,铁在不育系不同时期的花中积累,可能与CMS有关。研究[27]表明,Ca2+在雄性不育中具有重要的作用。光敏型雄性不育的水稻与小麦,小孢子晚期细胞质内Ca2+的异常积累可能导致了花粉的败育。在MRS通路中,酵母和哺乳动物等真核生物的正/负调控因子以及线粒体中的Ca2+参与其中[25]。与本研究中,2个不育系不同时期花/果以及2个时期花中钙含量高于保持系结果一致,钙含量升高可能是导致(gla)S和(sua)S不育的原因之一。水稻花粉小肽锌指蛋白(含Zn2+)基因OsFLZ13参与花药发育以及花粉育性的调控,其突变体植株结实率显著下降[28]。在拟南芥中,编码CCCH型锌指蛋白的基因At2g40140发生突变时,会导致小孢子仅剩下花粉壁的空壳[29]。在结球甘蓝中,编码的DHHC锌指蛋白的基因在保持系和细胞质雄性不育系植株中表现出巨大的差异,并在保持系中高表达[30]。在本研究中,(sua)S盛花期和青果期叶中锌含量低于另外2个基因型,可能与(sua)S种子产量有关。因活性氧大量积累导致花粉败育,在小麦[31]、棉花[32]及甜椒[33]中均有发现。锰超氧化物歧化酶(Mn-SOD)是一种重要的抗氧化剂,生理型雄性不育小麦的Mn-SOD基因在幼穗期、单核期和三核期表达量显著下调[34]。此外,锰同时是生长素吲哚乙酸氧化酶的辅助因子,影响内源激素水平,从而影响坐果率[35]。在本研究中,(sua)S不同发育时期和花蕾2个时期锰含量均显著低于(gla)S和MF K326,而(gla)S不同发育时期锰含量均最高,这种差异可能是导致雄性不育和(sua)S种子产量降低的原因之一。

4 结论

作为我国目前主栽的不育胞质类型,香甜烟草(N.suaveolens)胞质雄性不育系雌蕊畸形(花柱变扁或开裂),且比例很高(>50%),授粉后落花落果严重,种子产量极显著低于同型可育系MF K326和雌蕊正常的不育系(gla)S。通过系统研究不同时期和部位营养元素差异,发现(sua)S基因型导致雌雄蕊中钙和铁含量升高,磷酸盐、钾和锰含量降低,这一结果为可能的(sua)S雌蕊畸形改良和产量提升提供参考。

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中国果树, 1992(2):19-20.

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