转AtNHX1基因杨树Tr品系的耐盐性研究

doi:10.3773/j.issn.1005-264x.2010.05.010

摘要/Abstract

摘要：

以不同盐分强度处理欧美107杨(Populus × euramericana ‘Neva’) (Wt)和转拟南芥液泡膜Na⁺/H⁺逆向转运蛋白基因AtNHX1欧美107杨新品系(Tr)幼苗, 揭示Tr和Wt两品系幼苗耐盐性的差异, 探索拟南芥液泡膜Na⁺/H⁺逆向转运蛋白基因AtNHX1对提高杨树耐盐能力的效应。结果表明: 低盐处理下, Wt植株生长明显受到抑制, 其干重显著低于对照, 盐分强度加大后, 抑制作用更大, 其干重只有对照的50%; 而Tr植株在低盐处理下干重与对照差异不显著, 高盐处理时其干重为对照的74%。同时, 不同盐度处理下, Tr的干重均显著高于Wt, 且随着盐度升高, 两品系间植株干重差异增大。盐处理后, Tr植株叶片叶绿素和类胡萝卜素的含量均显著高于Wt, 并能维持较高的净光合速率(P_n)和PSII最大光化学效率(F_v/F_m); 在盐处理下虽然Tr叶片和根系均较Wt积累了更多的Na⁺, 但同时也维持了更高的K⁺和K⁺/Na⁺比率, 而且叶片对K⁺选择性的运输明显高于Wt; 同时, Tr叶片MDA含量和电解质渗漏率显著低于Wt。可见, 在盐处理下转AtNHX1植株较未转基因植株维持了更高的生长量、光合色素、光合能力和叶片质膜稳定性, 说明AtNHX1的转入能够显著提高欧美107杨的耐盐性。

关键词: AtNHX1, 离子区域化, 欧美杨, 耐盐性, 转基因植物

Abstract:

Aim Our objective was to test whether the overexpression of the Arabidopsis thaliana tonoplast Na⁺/H⁺ antiporter gene, AtNHX1, can improve salt tolerance in poplar.

Methods Transgenic and wild-type poplar (Populus × euramericana ‘Neva’) were subjected to low salt and high salt treatments. We investigated the effects of NaCl on the transgenic and wild-type poplar by measuring growth parameters, chlorophyll (Chl) and carotenoid (Car) content, net photosynthetic rate (P_n), maximal photochemical efficiency of PSII (F_v/F_m), ion content, leaf malondialdehyde (MDA) and electrolytic leakage after 30 days.

Important findings Compared with the control, the growth of wild-type plants was restrained significantly in the presence of both low salt and high salt. The dry weight of wild-type plants was significantly lower under salt stress than that of control, the dry weight decreased gradually with increasing NaCl concentration and their dry weight under high salt was just 50% of the control. However, the dry weight of transgenic plants in low salt was similar to the control up to the high salt treatment, where their dry weight was 74% of the control. Moreover, the dry weight of transgenic plants was significant higher than that of wild-type in both NaCl treatments, and the discrepancy of dry weight was increased with increasing NaCl concentration. In the presence of NaCl, Chl and Car content of transgenic plants were significantly higher than that of wild-type, and the transgenic plants maintained a remarkably high P_n and F_v/F_m. Although these transgenic plants accumulated more Na⁺ in their roots and leaf tissues under salinity conditions compared with the wild-type plants, they absorbed more K⁺and maintained a higher K⁺/Na⁺ratio. Moreover, these transgenic plants kept a lower MDA and electrolytic leakage level than that of wild-type. These findings indicate that transformation of AtNHX1 gene into poplar can confer plants more tolerance to salinity than wild-type poplar by maintaining better growth, higher Chl and Car content and improved P_n and F_v/F_m. These transgenic plants have potential for further applications in saline soil.

Key words: AtNHX1, ion compartmentation, Populus × euramericana ‘Neva’, salt tolerance, transgenic plant

姜超强, 郑青松, 刘兆普, 徐文君, 李洪燕, 李青. 转AtNHX1基因杨树Tr品系的耐盐性研究. 植物生态学报, 2010, 34(5): 563-570. DOI: 10.3773/j.issn.1005-264x.2010.05.010

JIANG Chao-Qiang, ZHENG Qing-Song, LIU Zhao-Pu, XU Wen-Jun, LI Hong-Yan, LI Qing. Salt tolerance of transgenic poplar by the introduction of AtNHX1 gene. Chinese Journal of Plant Ecology, 2010, 34(5): 563-570. DOI: 10.3773/j.issn.1005-264x.2010.05.010

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https://www.plant-ecology.com/CN/Y2010/V34/I5/563

图/表 7

表1 盐处理结束后栽培杨树土壤的可溶性盐含量(NaCl, %) (平均值±标准偏差)

Table 1 Soil soluble salt content of poplar’s pot after salt treatment (NaCl, %) (mean ± SD)

品系 Strain	对照 Control	低盐处理 Low salt treatment	高盐处理 High salt treatment
Tr	0.019 ± 0.004	0.395 ± 0.036	0.715 ± 0.035
Wt	0.019 ± 0.004	0.412 ± 0.023	0.745 ± 0.015

表2 盐处理30天转基因和未转基因植株干重的新复极差分析(平均值±标准偏差)

Table 2 Duncan test of dry weight of transgenic and wild-type poplar under different NaCl treatments for 30 days (mean ± SD)

处理 Treatment	基因型 Gene type	干重 Dry weight (g DW·plant^-1)	p < 0.05	p < 0.01
对照 Control	Tr	71.57 ± 2.74^a	a	A
对照 Control	Wt	70.08 ± 4.36^a	a	A
低盐 Low salt	Tr	65.44 ± 4.82^a	a	A
低盐 Low salt	Wt	51.58 ± 2.82^b	b	B
高盐 High salt	Tr	53.16 ± 2.38^b	b	B
高盐 High salt	Wt	35.15 ± 1.99^c	c	C

图1 盐处理对杨树叶片叶绿素(Chl)和类胡萝卜素(Car)含量的影响。 Wt、Tr, 同表1。柱状图中不同字母表示差异达5%显著水平。

Fig. 1 Effects of different salt treatments on chlorophyll (Chl) content and carotenoid (Car) content in leaves of poplar. Wt, Tr, see Table 1. Different letters in the histogram indicate significant difference at 5% level.

图2 盐处理对杨树叶片净光合速率(Pn)和最大光能效率(Fv/Fm)的影响。图注同图1。

Fig. 2 Effects of different salt treatments on net photosynthetic rate (Pn) and maximal photochemical efficiency (Fv/Fm) of poplar. Notes see Fig. 1.

表3 盐处理下欧美杨体内离子分布状况(平均值±标准偏差)

Table 3 Ion distribution in poplar under different salt treatments (mean ± SD)

处理 Treatment	基因型 Gene type	K⁺含量 K⁺ content (mmol·g^-1 DW)		Na⁺含量 Na⁺ content (mmol·g^-1DW)		K/Na比率 K/Na ratio
处理 Treatment	基因型 Gene type	根 Root	叶 Leaf	根 Root	叶 Leaf	根 Root	叶 Leaf
对照 Control	Tr	0.59 ± 0.05^a	0.81 ± 0.06^a	0.04 ± 0.00^e	0.04 ± 0.00^e	15.79 ± 2.06^a	21.11 ± 2.11^a
对照 Control	Wt	0.54 ± 0.06^ab	0.73 ± 0.07^ab	0.04 ± 0.00^e	0.04 ± 0.00^e	14.15 ± 1.57^b	19.88 ± 1.57^a
低盐 Low salt	Tr	0.55 ± 0.03^ab	0.77 ± 0.05^ab	0.38 ± 0.03^c	0.47 ± 0.01^c	1.44 ± 0.11^c	1.63 ± 0.08^b
低盐 Low salt	Wt	0.45 ± 0.04^cd	0.53 ± 0.05^c	0.33 ± 0.02^d	0.37 ± 0.01^d	1.35 ± 0.08^c	1.42 ± 0.15^b
高盐 High salt	Tr	0.51 ± 0.06^bc	0.70 ± 0.05^b	0.56 ± 0.03^a	0.79 ± 0.02^a	0.90 ± 0.09^c	0.88 ± 0.05^b
高盐 High salt	Wt	0.39 ± 0.04^d	0.40 ± 0.06^d	0.48 ± 0.03^b	0.57 ± 0.02^b	0.80 ± 0.12^c	0.71 ± 0.11^b

图3 盐处理对欧美杨K-Na选择性系数的影响。图注同图1。

Fig. 3 Effect of different salt treatments on the K-Na selectivity of poplar. Notes see Fig. 1.

图4 盐处理对杨树叶片MDA含量和电解质渗漏率的影响。图注同图1。

Fig. 4 Effects of different salt treatments on leaf MDA content and electrolytic leakage of poplar. Notes see Fig. 1.

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