外源硒预处理缓解茎瘤芥幼苗镉毒害的机制研究

doi:10.13560/j.cnki.biotech.bull.1985.2025-1079

摘要/Abstract

摘要：

目的为了明确外源硒（Se）预处理缓解茎瘤芥幼苗镉（Cd）毒害的机制，为今后合理使用Se从而保障茎瘤芥在Cd污染土壤的种植安全提供理论依据。方法以对Cd胁迫敏感的茎瘤芥主栽品种永安小叶为研究试材，通过水培试验，分别设置CK、CK+Se、Cd、Cd+Se 4种处理条件，研究外源Se对Cd胁迫下茎瘤芥幼苗Cd积累分配、光合同化能力、抗氧化能力等的影响。结果与CK相比，Cd胁迫处理显著抑制了茎瘤芥幼苗的生长，降低了植株叶绿素含量及光合效率，降低了抗氧化酶活性，提高了根系和叶片组织的Cd含量至CK的45.06和72.30倍。与单独Cd胁迫相比，外源Se预处理可明显促进Cd胁迫下茎瘤芥幼苗的生长，提高植株叶绿素含量、Pn、Fv/Fm、ΦPSⅡ、ETR和qP值，提高SOD和CAT活性，增加ASA、GSH、总酚、螯合肽和半胱氨酸含量，并下调Cd吸收和转运相关基因IRT1、Nramp1、HMA2、HMA3的表达量至单独Cd胁迫处理的46.85%‒70.33%，减少根、叶组织Cd的积累量并提高细胞壁的Cd含量占比。结论 Cd胁迫条件下，外源Se主要通过提高光合系统的转化效率、提高抗氧化能力、增强对Cd的螯合能力及区隔化作用，以及下调Cd吸收和转运相关基因的表达水平，从而缓解Cd对茎瘤芥幼苗产生的毒害作用。

关键词: 茎瘤芥, 镉胁迫, 硒, 镉毒害, 抗氧化能力

Abstract:

Objective To clarify the mechanism of exogenous selenium (Se) pretreatment alleviating cadmium (Cd) toxicity in Brassica juncea var. tumida Tsen et Lee seedlings, and to provide a theoretical basis for the rational application of Se to ensure the safe cultivation of Brassica juncea var. tumida Tsen et Lee in Cd-contaminated soils. Method The dominant cultivar ‘Yonganxiaoye’ which is sensitive to Cd stress was used as the experimental material, and a hydroponic experiment was conducted with four treatments: CK, CK+Se, Cd, and Cd+Se, thus to study the effects of exogenous selenium on Cd accumulation and distribution, photosynthetic assimilation capacity, and antioxidant capacity of B. juncea var. tumida Tsen et Lee seedlings under Cd stress. Results Compared with CK, Cd stress treatment markedly inhibited the growth of B. juncea var. tumida Tsen et Lee seedlings, reduced the chlorophyll content and photosynthetic efficiency of the plants, decreased the antioxidant enzymes activities, and increased the Cd content in both root and leaf tissues to 45.06 and 72.30 times that of CK, respectively. Compared with Cd stress alone, exogenous Se pretreatment significantly promoted the growth of B. juncea var. tumida Tsen et Lee seedlings under Cd stress, increased the chlorophyll content, Pn, Fv/Fm, ΦPSII, ETR, and qP values of the plants, enhanced the activities of SOD and CAT, and increased the contents of ASA, GSH, total phenol, phytochelatin, and cysteine, down-regulated the expression levels of Cd uptake and transport-related genes (IRT1, Nramp1, HMA2, and HMA3) to 46.85%‒70.33% of those treated by Cd stress alone, reduced Cd accumulation in both root and leaf tissues and increased the proportion of Cd content in the cytoderm. Conclusion It can be seen that exogenous Se alleviates Cd toxicity in B. juncea var. tumida Tsen et Lee seedlings under Cd stress, mainly by improving the conversion efficiency of photosynthetic system, enhancing the antioxidant capacity,^{strengthening the Cd chelating capacity and compartmentalization, and down-regulating the Cd uptake and transport-related genes expression levels.}

Key words: Brassica juncea var. tumida Tsen et Lee, cadmium stress, selenium, cadmium toxicity, antioxidant capacity

莫言玲, 刘源橼, 裴佳玲, 樊襄曼, 邓兴连, 曾静, 刘义华. 外源硒预处理缓解茎瘤芥幼苗镉毒害的机制研究[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1079.

MO Yan-ling, LIU Yuan-yuan, PEI Jia-ling, FAN Xiang-man, DENG Xing-lian, ZENG Jing, LIU Yi-hua. Study on the Mechanism of Exogenous Selenium Pretreatment Alleviating Cadmium Toxicity in Brassica juncea var. tumida Tsen et Lee Seedlings[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1079.

图/表 9

表1 用于实时荧光定量分析的引物序列

Table 1 Primer sequences for quantitative real-time PCR analysis

基因 Gene	基因登录号 Gene accession number	正向引物 Forward primer sequence （5′‒3′）	反向引物 Reverse primer （5′‒3′）
Actin3	BjuA014894	GGCTACTCTTTCACCACGAC	GGATACCAGCATTCTCCATAC
IRT1	AY087095.1	TGGCATTCTTTTTCGCGGTG	GCCGAGCATGCATTGAGAAG
IRT2	BT025714.1	CTCGTCGACCTTCTGGCTAC	ACTTGGCGACGACAGACATT
Nramp1	AF165125.1	CCCCGAAGACCGTGCTAAAT	TACCCACCACGTTTCGTAGC
HMA2	NM119157.3	GAGGATGCCACATGGTTGGA	CTTTGGTACGGCGGAAGAGT
HMA3	DQ446885.1	AACCTCGACGCTATGCACAA	GCTTGCCACGTCATCATTGG
HMA4	BjuO003139	TTCCCCACAAGAATCGCTCC	CACTCGAACCTTCCACGTCA

图1 各处理条件下茎瘤芥幼苗植株的外部形态特征CK：霍格兰全营养液；CK+Se：霍格兰全营养液+硒预处理；Cd：镉胁迫处理；Cd+Se：镉胁迫处理+硒预处理。bar=1 cm。下同

Fig. 1 External morphology of Brassica juncea var. tumida Tsen et Lee seedlings under different treatment conditionsCK: Hoagland’s complete nutrient solution. CK+Se: Hoagland’s complete nutrient solution + Selenium pretreatment. Cd: Cadmium stress treatment. Cd+Se: Cadmium stress treatment + Selenium pretreatment. bar=1 cm. The same below

表2 外源硒预处理对镉胁迫下茎瘤芥幼苗生长的影响

Table 2 Effect of exogenous selenium pretreatment on the growth of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

处理 Treatment	叶片数 Leaf number	株高 Plant height （cm）	根长 Root length （cm）	地上鲜重 Shoot fresh weight （g）	地下鲜重 Root fresh weight （g）	地上干重 Shoot dry weight （g）	地下干重 Root dry weight （g）
CK	6.56±0.53a	7.06±0.29a	19.19±0.45a	7.52±0.34a	0.64±0.05a	0.64±0.08a	0.05±0.01ab
CK+Se	6.67±0.50a	7.12±0.36a	19.79±1.53a	7.55±0.24a	0.65±0.04a	0.63±0.08a	0.05±0.00a
Cd	5.89±0.33b	4.22±0.30c	12.78±1.65b	4.37±0.26c	0.35±0.03c	0.25±0.02c	0.04±0.00c
Cd+Se	6.44±0.53a	6.26±0.31b	18.64±0.76a	6.53±0.29b	0.51±0.04b	0.36±0.02b	0.04±0.00bc

图2 外源硒预处理对镉胁迫下茎瘤芥幼苗叶绿素含量、光合参数和叶绿素荧光参数的影响

Fig. 2 Effects of exogenous selenium pretreatment on the chlorophyll content, photosynthetic parameters, and chlorophyll fluorescence parameters of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

图3 外源硒预处理对镉胁迫下茎瘤芥幼苗MDA、H2O2和O2-含量的影响

Fig. 3 Effects of exogenous selenium pretreatment on the MDA, H2O2, and O2-contents of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

图4 外源硒预处理对镉胁迫下茎瘤芥幼苗抗氧化酶活性和抗氧化物质含量的影响

Fig. 4 Effects of exogenous selenium pretreatment on the antioxidant enzyme activities and antioxidant contents of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

图5 外源硒预处理对镉胁迫下茎瘤芥幼苗螯合肽和半胱氨酸含量的影响

Fig. 5 Effects of exogenous selenium pretreatment on the chelating peptide and cysteinet contents of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

图6 外源硒预处理对镉胁迫下茎瘤芥幼苗镉积累量的影响

Fig. 6 Effects of exogenous selenium pretreatment on the cadmium accumulations of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

图7 外源硒预处理对镉胁迫下茎瘤芥幼苗根系镉吸收和转运相关基因表达量的影响

Fig. 7 Effects of exogenous selenium pretreatment on the expressions of genes related to cadmium uptake and transport in the roots of B. juncea var. tumida Tsen et Lee seedlings under cadmium stress

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