东京四照花钙组分与渗透调节物质对盐胁迫的响应

doi:10.16258/j.cnki.1674-5906.2023.04.006

生态环境学报 ›› 2023, Vol. 32 ›› Issue (4): 687-696.DOI: 10.16258/j.cnki.1674-5906.2023.04.006

东京四照花钙组分与渗透调节物质对盐胁迫的响应

袁佳秋¹^,²(), 孙大伟², 杨玲³, 洑香香²^,^*()

1.江苏食品药品职业技术学院，江苏淮安 223005
2.南方现代林业协同创新中心/南京林业大学林学院，江苏南京 210037
3.上海市野生动植物和自然保护地研究中心，上海 200336

收稿日期:2023-03-03 出版日期:2023-04-18 发布日期:2023-07-12
通讯作者: *E-mail: xxfu@njfu.edu.cn
作者简介:袁佳秋（1994年生），女，讲师，博士研究生，主要从事抗逆机制研究。E-mail: 20221026@jsfpc.edu.cn
基金资助:
江苏省林业科技创新与推广项目(LYKJ[2018]06);江苏省高校重点学科建设项目(PAPD);江苏省研究生科研实践与创新计划项目(KYCX21_0888)

Responses of Calcium Composition and Osmotica under Salt Stress in Cornus hongkongensis Subsp. Tonkinensis (W. P. Fang) Q. Y. Xiang

YUAN Jiaqiu¹^,²(), SUN Dawei², YANG Ling³, FU Xiangxiang²^,^*()

1. Jiangsu Food &Pharmaceutical Science College, Huai’an 223005, P. R. China
2. Co-Innovation Center for the Sustainable Forestry in Southern China/College of forestry, Nanjing forestry university, Nanjing 210037, P. R. China
3. Shanghai Wildlife and Protected Natural Areas Research Center, Shanghai 200336, P. R. China;

Received:2023-03-03 Online:2023-04-18 Published:2023-07-12

摘要/Abstract

摘要：

随着滨海地区土壤盐渍化程度不断扩大，选育耐盐性和观赏价值并存的木本植物进行引种栽培，将有利于盐渍土壤资源的利用和生态系统的改善。东京四照花（Cornus hongkongensis subsp. tonkinensis）作为观赏型苗木，从幼苗的钙组分与渗透物质方面探讨其盐胁迫响应机制，为其在滨海地区推广种植提供理论依据。设置4种质量分数梯度的盐浓度：0（CK）、0.20%（T1）、0.30%（T2）和0.45%（T3），水培法培养幼苗。盐处理幼苗的盐害等级（I_SD）随着盐分浓度的升高和胁迫时间的延长而增加。盐胁迫5 d时，各处理组的幼苗I_SD和成活率未有明显变化；盐胁迫30 d时，T1、T2和T3处理组的I_SD分别为47.80%、52.00%和71.20%，对应的成活率分别为82.60%、84.00%和56.00%。胁迫初期（5 d），T1、T2和T3保卫细胞中Ca²⁺浓度较CK分别增加了13.98%、22.54%和17.33%。同时，T1、T2和T3的气孔孔径的宽长比（W/L）比CK分别减少了23.60%、25.98%和36.05%。其中，仅T3处理保卫细胞中Ca²⁺浓度随胁迫时间延长而显著升高。胁迫至30 d，T3处理下的水溶性钙（H₂O-Ca）、草酸钙（HCl-Ca）、果胶酸钙（NaCl-Ca）质量分数分别降低至4.99、7.12、4.04 mg?g^?1。同时，随着盐浓度的升高，草酸钙晶体在叶肉细胞中破碎化程度明显。与CK相比，T1、T2和T3草酸钙晶体在叶肉细胞中面积占比分别显著降低了43.10%、15.25%、69.90%。T3处理下的可溶性蛋白（SP）、可溶性糖（SS）和脯氨酸（Pro）含量较CK分别增加了20.04 %、15.38%和19.74%。东京四照花短期（30 d）可承受0.45%的盐胁迫。幼苗通过提高Ca²⁺浓度，促进可溶性蛋白的合成来提高其耐盐性。此外，草酸钙降解释放出的Ca²⁺可能是缓解盐胁迫对东京四照花幼苗损伤的有效策略。

关键词: 盐胁迫, 钙组分, 草酸钙, 渗透物质

Abstract:

The continuous expansion of soil salinization in coastal areas necessitates the selection and breeding of woody plants with both salt tolerance and ornamental value. This practice is vital for utilizing saline soil resources and enhancing the ecosystem. To elucidate the response mechanism to salt stress, we analyzed the effects of varying concentrations of salt stress on calcium components and osmotic substances in Cornus hongkongensis subsp. tonkinensis seedlings. The results can serve as a theoretical foundation for promoting and cultivating ornamental dogwood in coastal areas. The study involved the hydroponic cultivation of seedlings with four gradient concentrations of sea salt [0 (CK), 0.20% (T1), 0.30% (T2), and 0.45% (T3)]. The degree of salt damage index (ISD) in salinized seedlings increased with both the rising salt concentration and the extension of stress duration. After 5 days of salt stress, the ISD and survival rate in salinized seedlings showed no significance. Salinized seedlings exposed to T1, T2, and T3 exhibited ISD of 47.80%, 52.00%, and 71.20% after 30 days of salt stress, and the corresponding survival rates were 82.60%, 84.00%, and 56.00%. During short- term saline stress (5 d), the Ca²⁺ concentration in the guard cells of leaves exposed to T1, T2, and T3 increased, reaching 13.98%, 22.54%, and 17.33% above CK, separately. Simultaneously, the width-to-length ratios of pore in T1, T2, and T3 were lower than that in CK by 23.60%, 25.98%, and 36.05%, respectively. Thereof, the Ca²⁺ concentration in the guard cells of leaves exposed to T3 increased strikingly with the extension of stress duration. After 30 days of salt stress, the mass fractions of water-soluble calcium, oxalate calcium, and pectin-bound calcium in T3 significantly decreased to the values of 4.99, 7.12, and 4.04 mg·g^?1, respectively. Additionally, as the salt concentration increased, there was obvious fragmentation of calcium oxalate crystals in mesophyll cells. The area proportion of calcium oxalate crystals in mesophyll cells exposed to T1, T2, and T3 fell by 43.10%, 15.25%, and 69.90% above CK, separately. The content of soluble protein (SP), soluble sugar (SS), and proline (Pro) in the leaves of T3 treatment increased by 20.04 %, 15.38%, and 19.74% compared to CK, respectively. The results indicate that C. hongkongensis subsp. tonkinensis exhibits tolerance to 0.45% salt stress for 30 days. The salt tolerance of C. hongkongensis subsp. tonkinensis seedlings are believed to be achieved through an increase in both the Ca²⁺ content and soluble protein content. In addition, the release of Ca²⁺ through the decomposition of calcium oxalate may be an effective strategy to mitigate the damage caused by salt stress in dogwoods.

Key words: salt stress, calcium component, calcium oxalate, osmotic substances

中图分类号:

Q945
X17

袁佳秋, 孙大伟, 杨玲, 洑香香. 东京四照花钙组分与渗透调节物质对盐胁迫的响应[J]. 生态环境学报, 2023, 32(4): 687-696.

YUAN Jiaqiu, SUN Dawei, YANG Ling, FU Xiangxiang. Responses of Calcium Composition and Osmotica under Salt Stress in Cornus hongkongensis Subsp. Tonkinensis (W. P. Fang) Q. Y. Xiang[J]. Ecology and Environment, 2023, 32(4): 687-696.

图/表 11

图1 盐胁迫30 d下东京四照花叶片盐损伤症状

Figure 1 Salt stress damage symptoms in leaves of C. hongkongensis subsp. tonkinensis for 30 days

图2 不同盐浓度下东京四照花盐损伤指数和成活率方块代表第五天的数据，三角代表第30天的数据；实线代表盐损伤指数，虚线代表成活率。CK、T1、T2、T3分别表示4种质量分数的海盐浓度，依次为0%、0.20%、0.30%、0.45%。下同

Figure 2 Salt damage index (ISD) and survival rate of C. hongkongensis subsp. tonkinensis under different salt concentrations

图3 盐胁迫下东京四照花叶片保卫细胞Ca2+分布用荧光显微镜观察对照和盐处理后荧光Fluo-3AM染色叶中的Ca2+。（a和i）CK处理、（b和j）T1处理、（c和k）T2处理和（d和l）T3处理的叶片下表皮图像，（e、f、g和h）和（m、n、o和p）分别为盐处理5 d和30 d时红色框内对应的气孔放大图，所有处理中均可见Ca2+在保卫细胞周围的定位。标尺为50 μm

Figure 3 Ca2+ distribution in guard cells of leaves in C. hongkongensis subsp. tonkinensis under salt stress

图4 盐胁迫下东京四照花叶片保卫细胞Ca2+荧光强度变化图中数据为平均值±标准误，n=3。不同大写字母表示不同处理同一时间的多重比较（P<0.05）；根据独立样本T检验，*和**表示同一处理不同时间的差异显著（P<0.05，0.01）。下同

Figure 4 Change of fluorescence intensity of Ca2+ in guard cells of leaves in C. hongkongensis subsp. tonkinensis under salt stress

表1 盐胁迫对东京四照花叶片气孔相关参数的影响

Table 1 Effect of salt stress on stoma of leaves in C. hongkongensis subsp. tonkinensis

处理	孔隙长度/µm			孔隙宽度/µm			宽长比W/L
处理	5 d	30 d	Sig.	5 d	30 d	Sig.	5 d	30 d	Sig.
CK	7.51±0.27A	8.53±0.08A	*	2.86±0.33A	3.05±0.03A	ns	0.38±0.03A	0.36±0.00A	ns
T1	4.79±0.80B	4.78±0.53C	ns	1.38±0.19B	1.33±0.17C	ns	0.29±0.03B	0.27±0.01C	ns
T2	5.16±0.50B	5.01±0.13C	ns	1.43±0.15B	1.37±0.02C	ns	0.28±0.01B	0.28±0.01C	ns
T3	4.65±0.84B	7.08±0.72B	**	1.16±0.32B	2.04±0.17B	*	0.24±0.03B	0.29±0.01B	ns

图5 盐胁迫对东京四照花叶片气孔形态的影响图a－d分别为在盐胁迫5 d时，CK、T1、T2、T3组叶片气孔电镜图；图e－h分别为在盐胁迫30 d时，CK、T1、T2、T3组叶片气孔电镜图；图中标尺为40 μm

Figure 5 Effect of salt stress on stoma morphology of leaves in C. hongkongensis subsp. tonkinensis

图6 不同盐浓度对东京四照花叶片不同形态钙质量分数的影响

Figure 6 Effect of salt stress on mass fraction of different calcium forms in leaves of C. hongkongensis subsp. tonkinensis

图7 盐胁迫下东京四照花叶片草酸钙晶体显微观察图a－d分别为在盐胁迫5 d时，CK、T1、T2、T3组叶片草酸钙晶体显微图；图e－h分别为在盐胁迫30 d时，CK、T1、T2、T3组叶片草酸钙晶体显微图；图中标尺为100 μm

Figure 7 Microscopic observation on calcium oxalate crystals in salt-stressed leaves of C. hongkongensis subsp. tonkinensis

图8 盐胁迫下东京四照花叶片草酸钙晶体的比面积

Figure 8 Area ratio of calcium oxalate crystal in leaves in C. hongkongensis subsp. tonkinensis under salt stress

表2 东京四照花盐胁迫下渗透调节物质响应

Table 2 Response of osmotic regulators in the salt-stressed seedlings of C. hongkongensis subsp. tonkinensis

指标	处理	胁迫时间/d/
指标	处理	5 d	30 d	Sig.
可溶性蛋白质量分数/ (mg∙g⁻¹)	CK	5.39±0.40A	5.54±0.35B	ns
	T1	5.17±0.48A	5.44±0.12B	ns
	T2	5.95±0.38A	5.60±0.43B	ns
	T3	5.81±0.52A	6.65±0.45A	ns
可溶性糖质量分数/ (mg∙g⁻¹)	CK	21.91±1.87A	19.77±1.83B	ns
	T1	24.60±1.52A	20.29±2.12B	*
	T2	21.72±2.22A	25.13±1.70A	ns
	T3	22.50±2.17A	22.81±1.80AB	ns
脯氨酸质量分数/ (μg∙g⁻¹)	CK	1436.83±256.96A	1221.83±72.96AB	ns
	T1	1089.86±66.99B	897.59±196.75B	ns
	T2	971.23±111.53B	956.41±214.04B	ns
	T3	1042.41±229.38B	1463.03±244.93A	ns

表3 叶片钙组分和渗透调节物质的相关系数（r）

Table 3 Correlation coefficients among leaves calcium component and osmoregulation substances

指标	I_SD	Ca²⁺ FI	W/L	SP	SS	Pro	H₂O-Ca	NaCl-Ca	HAc-Ca	HCl-Ca	CaOx AR
I_SD	1
Ca²⁺ FI	0.628**	1
W/L	−0.305	−0.221	1
SP	0.606**	0.554**	−0.237	1
SS	0.131	0.139	−0.355	−0.001	1
Pro	0.137	0.072	0.512*	0.265	−0.047	1
H₂O-Ca	−0.915**	−0.692**	0.351	−0.515**	−0.070	0.056	1
NaCl-Ca	−0.865**	−0.478*	0.177	−0.644**	0.129	−0.209	0.777**	1
HAc-Ca	−0.209	−0.065	0.091	−0.214	0.120	0.134	0.249	0.288	1
HCl-Ca	−0.819**	−0.453*	0.211	−0.480*	0.060	0.133	0.846**	0.764**	0.498*	1
CaOx AR	−0.784**	−0.543**	0.322	−0.731**	0.118	−0.122	0.734**	0.747**	0.266	0.796**	1

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东京四照花钙组分与渗透调节物质对盐胁迫的响应

Responses of Calcium Composition and Osmotica under Salt Stress in Cornus hongkongensis Subsp. Tonkinensis (W. P. Fang) Q. Y. Xiang

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