盐碱胁迫对甘蓝型油菜生理及分子机制的影响

doi:10.7668/hbnxb.20192954

摘要/Abstract

摘要：

为探究盐碱胁迫对甘蓝型油菜的生理及分子机制的影响,以甘蓝型油菜华油杂62为试验材料,对油菜种子及幼苗进行不同浓度的复合盐、复合碱及复合盐碱溶液处理,测定种子的发芽率以及甘蓝型油菜叶片中叶绿素含量、脯氨酸含量、可溶性糖含量、抗氧化酶活性等生理指标,以高效液相色谱法测定油菜叶片中甜菜碱积累量,利用qRT-PCR 技术对甜菜碱合成途径中的关键酶基因胆碱单加氧酶基因(CMO)进行分析。结果表明,人工模拟的不同浓度盐碱溶液中,对种子萌发的伤害程度大小表现为复合盐碱>碱>盐;低浓度盐碱溶液促进油菜叶片叶绿素形成,高浓度盐碱溶液抑制叶绿素形成;盐碱胁迫显著提高了脯氨酸与可溶性糖含量,高盐碱溶液(YJ75,含盐碱75 mmol/L)处理21 d,脯氨酸与可溶性糖含量分别为对照组的65.99,5.21倍;盐碱胁迫增加了丙二醛的含量;盐碱胁迫显著提高了过氧化物酶(POD)活性,与对照组相比,高盐碱(YJ75)溶液处理21 d后的POD含量提高了2.26倍,在复合盐与复合碱处理第14 天,POD含量达到最高值,而超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性变化规律不明显,且在盐胁迫过程中发挥作用较低;盐碱胁迫显著提高了关键酶基因CMO的表达量,从而调控甜菜碱的积累量增多。综上,盐碱胁迫对甘蓝型油菜的伤害程度大小表现为复合盐碱>碱>盐,在高盐碱胁迫下,甘蓝型油菜会大量积累甜菜碱以降低自身所受的伤害。

关键词: 甘蓝型油菜, 盐碱胁迫, 生理指标, 高效液相色谱法, 甜菜碱, qRT-PCR

Abstract:

In order to explore the effects of saline-alkali stress on the physiological and molecular mechanism of Brassica napus L.,using Huayouza 62 as experimental material.The seedlings of Brassica napus L.were treated with different concentrations of compound salt,compound alkali and compound saline-alkali solution.Physiological indexes such as seed germination rate,chlorophyll content,proline content,soluble sugar content and antioxidant enzyme activity in Brassica napus L.leaves were determined.The accumulation of betaine in rape leaves was determined by high performance liquid chromatography(HPLC).The key enzyme gene choline monooxygenase gene(CMO)in betaine synthesis pathway was analyzed by qRT-PCR technique.The results showed that the degree of damage to seed germination in artificially simulated saline-alkali solution of different concentrations was compound saline-alkali>alkali>salt.Low concentration saline-alkali solution promoted chlorophyll formation in rape leaves,while high concentration saline-alkali solution inhibited chlorophyll formation,saline-alkali stress significantly increased the contents of proline and soluble sugar,and the contents of proline and soluble sugar in high saline-alkali solution(YJ75,saline-alkali 75 mmol/L)for 21 d were 65.99 and 5.21 times higher than those in the control group,respectively,and the content of malondialdehyde was increased by saline-alkali stress.Saline-alkali stress significantly increased the activity of peroxidase(POD).Compared with the control group,the content of POD in high saline-alkali solution(YJ75)increased by 2.26 times after 21 d,and the content of POD reached the highest value on the 14th day after treatment with compound salt and compound alkali,however, the activity changes of superoxide dismutase (SOD) and catalase (CAT) were not obvious, and the role of species in the process of saline-alkali stress was low.Saline-alkali stress significantly increased the expression of key enzyme gene CMO,thus regulating the accumulation of betaine.In summary,the damage degree of saline-alkali stress to Brassica napus L. was compound saline-alkali > alkali > salt.Under high saline-alkali stress,Brassica napus would accumulate a large amount of betaine to reduce the damage.

Key words: Brassica napus L., Saline-alkali stress, Physiological indexes, HPLC, Betaine, qRT-PCR

李班, 吕莹, 杨明煊, 宋婷, 于放, 刘志文. 盐碱胁迫对甘蓝型油菜生理及分子机制的影响[J]. 华北农学报, 2022, 37(3): 86-93. doi: 10.7668/hbnxb.20192954.

LI Ban, LÜ Ying, YANG Mingxuan, SONG Ting, YU Fang, LIU Zhiwen. Effects of Saline-alkali Stress on Physiology and Molecular Mechanism of Brassica napus L.[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 86-93. doi: 10.7668/hbnxb.20192954.

http://www.hbnxb.net/CN/Y2022/V37/I3/86

图/表 13

表1 引物序列

Tab.1 Primer sequences

引物名称 Primer name	引物序列( 5'-3') Primer sequence( 5'-3')
PP2A-F1602	GTCAACAATCCGCACTACCTA
PP2A-R1711	ACCACGACGGGAAGAAAC
CMO-F674	CTGTATGGGGACCTTTTGTT
CMO-R799	CGCTCAACCTCCCTACAGA

表2 盐碱胁迫对种子发芽率的影响

Tab.2 The effect of saline-alkali stress on the seed germination

处理 Management		含盐量/(mmol/L) Salt content	发芽率/% Germination rate
CK	H₂O	0	100±0.00a
复合盐	Y25	25	98±0.94a
Compound	Y50	50	96±1.63a
salt	Y75	75	90±0.81b
复合碱	J25	25	68±2.44c
Compound	J50	50	18±2.16e
alkali	J75	75	4±1.41f
复合盐碱	YJ25	25	48±3.55d
Compound	YJ50	50	0±0.00g
saline-alkali	YJ75	75	0±0.00g

图1 复合盐、复合碱、复合盐碱对甘蓝型油菜叶片叶绿素的影响不同小写字母表示差异显著(P<0.05)。图2-11同。

Fig.1 Effect of compound salt,compound alkali,compound saline-alkali on chlorophyll in leaves of Brassica napus L. Different lowercase letters indicate significant difference(P<0.05).The same as Fig.2-11.

图2 复合盐、复合碱、复合盐碱对甘蓝型油菜叶片脯氨酸含量的影响

Fig.2 Effect of compound salt,compound alkali,compound saline-alkali on the proline content in leaves of Brassica napus L.

图3 复合盐、复合碱、复合盐碱对甘蓝型油菜叶片可溶性糖含量的影响

Fig.3 Effect of compound salt,compound alkali,compound saline-alkali on the soluble sugar content in leaves of Brassica napus L.

图4 复合盐、复合碱、复合盐碱对甘蓝型油菜叶片丙二醛含量的影响

Fig.4 Effect of compound salt,compound alkali,compound saline-alkali on malondialdehyde content in leaves of Brassica napus L.

图5 盐胁迫对甘蓝型油菜叶片抗氧化酶活性的影响

Fig.5 Effect of salt stress on antioxidant enzyme activity in leaves of Brassica napus L.

图6 碱胁迫对甘蓝型油菜叶片抗氧化酶活性的影响

Fig.6 Effect of alkali stress on antioxidant enzyme activity in leaves of Brassica napus L.

图7 盐碱胁迫对甘蓝型油菜叶片抗氧化酶活性的影响

Fig.7 Effect of saline-alkali stress on antioxidant enzyme activity in leaves of Brassica napus L.

图8 盐胁迫对合成甜菜碱关键酶基因相对表达量的影响

Fig.8 Effect of salt stress on relative expression of key enzyme genes in betaine synthesis

图9 碱胁迫对合成甜菜碱关键酶基因相对表达量的影响

Fig.9 Effect of alkali stress on relative expression of key enzyme genes in betaine synthesis

图10 盐碱胁迫对合成甜菜碱关键酶基因相对表达量的影响

Fig.10 Effect of saline-alkali stress on relative expression of key enzyme genes in betaine synthesis

图11 甘蓝型油菜叶片中甜菜碱含量

Fig.11 The betaine content in leaves of Brassica napus L.

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