四种暖季型草坪草抗盐生理及根系蛋白表达差异研究
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摘要
盐胁迫可造成草坪草在生长、生理生化和蛋白等水平上的变化，是限制草坪草生长的主要环境因子之一。研究草坪草抗盐机理以及草坪草对盐的适应和调节机制，对于筛选适合不同土壤盐浓度的草坪草种具有重要的意义。本文以华东地区常用的四种暖季型草坪草为材料，进行了抗盐性评价与筛选，确定了海滨雀稗、狗牙根、结缕草和假俭草四个暖季型草坪草种的耐盐阈值；对这四种暖季型草坪草的泌盐现象及泌盐能力进行了初步研究；选择抗盐性强的海滨雀稗和盐敏感的假俭草为研究对象，比较了其在盐胁迫下光合生理的差异，并通过双向凝胶电泳分析了盐胁迫下根系蛋白的表达差异。主要研究结果如下：
1．暖季型草坪草的耐盐阈值
以杂交狗牙根‘Tifway’(Cynodon dactylon×C. transvaalensis)、假俭草‘Civil’(Eremochloa ophiuroides)、海滨雀稗‘Salam’(Paspalum vaginatum)和结缕草‘Matrella’(Zoysia matrella L.Merr)四种暖季型草坪草为材料，研究了不同盐(NaCl)浓度处理对草坪草生长、生理的影响。结果表明：海滨雀稗耐盐性最好，海滨雀稗的目测质量、生长量、叶片相对含水量、叶绿素指数、光化学效率在600mM盐浓度处理36d时分别下降了52.2%、75.9%、62.1%、59.4%、72.9%，相对电导率上升至71.5%，与其他三个草种相比，海滨雀稗各指标仍能维持较高的水平，膜伤害程度相对较小；结缕草耐盐性次之，600mM盐处理36d时目测质量、生长量、叶片相对含水量、叶绿素指数、光化学效率分别下降了85.2%、68.8%、70.5%、73.9%、49.3%，相对电导率上升至81.8%，表现出较好的耐盐性；耐盐性表现较差的为狗牙根和假俭草，在600mM盐浓度处理36d时，两种草的各指标皆下降至0，膜伤害程度达到最大，植物已枯死。以36d时的各生理指标与盐处理浓度建立回归方程，以最小显著差异性检验(LSD0.05)的置信区间分别求出各指标与对照无显著差异的盐浓度临界值。选取最低的临界盐浓度作为该植物的耐盐阈值。求得四个草种的耐盐阈值分别为为：海滨雀稗261mM、结缕草203mM、狗牙根135mM、假俭草131mM。
2．暖季型草坪草的泌盐能力
对杂交狗牙根‘Tifway’(Cynodon dactylon×C. transvaalensis)、假俭草‘Civil’(Eremochloa ophiuroides)、海滨雀稗‘Salam’(Paspalum vaginatum)和上海结缕草‘JD-1’(Zoysia japomca)叶表面盐分测定及活体显微观察发现，最耐盐的海滨雀稗和盐敏感的假俭草没有泌盐功能，而结缕草和狗牙根则具有泌盐功能。通过荧光反射显微镜观察发现，结缕草和狗牙根泌盐结构为盐腺，且狗牙根泌盐量比结缕草要高，在盐处理的20d狗牙根泌盐量为结缕草的1.6倍。主要原因为狗牙根盐腺密度比结缕草大，结缕草叶片盐腺数量平均为66个/mm~2，而狗牙根叶片盐腺数量为169个/mm~2，是结缕草盐腺数量的2.6倍。通过荧光反射显微观察，狗牙根和结缕草的泌盐随着盐处理时间的延长，泌盐量不断增加。但两者盐腺数量在盐胁迫下没有明显变化。狗牙根的泌盐能力高于结缕草，但其抗盐能力却低于结缕草，且耐盐能力最强的海滨雀稗和盐敏感的假俭草中均未发现具有泌盐能力，说明泌盐机制及泌盐能力的大小并不直接决定暖季型草坪草的抗盐能力。
3．海滨雀稗和假俭草对盐胁迫的光合响应
以耐盐的海滨雀稗和盐敏感的假俭草为材料，利用盆栽研究不同浓度盐处理(0,300和500mM NaCl)下的光合生理响应变化。海滨雀稗和假俭草的草坪目测质量、叶片相对含水量及Fv/Fm均随处理时间及处理浓度的递增而呈下降趋势，叶片质膜透性呈上升趋势；盐敏感的假俭草的各指标的变化均明显大于耐盐的海滨雀稗。盐胁迫下两个草种表现出不同的光合响应变化：海滨雀稗在低浓度盐处理初期，光合速率的下降主要是由气孔性因素即气孔关闭引起的胞间CO2浓度降低所造成的，随着处理时间的延长，海滨雀稗光合速率有所恢复，此时海滨雀稗gs有所回升，Ci与对照无显著差异，叶绿素含量、Fv/Fm及Rubisco大亚基均未显著下降。假俭草在两浓度处理下光合速率的下降主要是由非气孔性因素作用的结果，假俭草的光合速率及各生理指标持续下降，Ci持续升高，Rubisco大亚基含量显著下降。海滨雀稗比假俭草抗盐是由于海滨雀稗具有较强的光合恢复能力。
4．海滨雀稗和假俭草根系蛋白表达差异
以抗盐海滨雀稗和盐敏感假俭草为材料，利用2-DE研究两者盐胁迫下根系蛋白差异表达的情况。结果表明，根系渗透势和根系活力在盐胁迫下随着处理时间的延长而不断下降，根系质膜透性随处理时间的延长不断上升。但假俭草各生理指标的变化程度要比海滨雀稗严重。2-DE检测结果表明，海滨雀稗中有8个蛋白点显著上调，14个蛋白点显著降解；假俭草中分别有19和16个蛋白点显著上调和降解。对海滨雀稗8个上调蛋白进行质谱鉴定，7个蛋白点鉴定成功，鉴定结果为：两个蛋白为ATP合成酶链(ATPS链)，五个为盐胁迫响应蛋白，分别是：过氧化物酶(POD，2.36倍)，苹果酸脱氢酶(cMDH，5.84倍)，抗坏血酸过氧化物酶(APX，4.03倍)及线粒体ATP合成酶链(ATPS链，2.26倍和4.78)；2个为新蛋白：hypothetical protein LOC100274119(5.01倍)和FlavoproteinwrbA (2.20倍)。通过对POD和ATP合成酶链进行蛋白免疫印迹验证发现，海滨雀稗在盐处理下，POD和ATPS链的表达明显高于对照，而假俭草对照与处理间无显著差异。本研究结果表明，和假俭草相比，海滨雀稗较高的抗盐性可能与它较高的POD、APX活性及较高的ATP合成能力有关。
Salinity is one of the major abiotic factors limiting plant growth and inducesvarious changes in the growth, physiological and proteomic levels. Understanding themechanisms of salinity resistance and adaptation in turfgrass will provide the crucialreference to select suitable turfgrass variety for salinity soils and imformation forsalinity-tolerant variety development. In this study, existing turfgrass varieties werescreened for the salinity tolerance and the salt tolerance thresholds of fourwarm-season turfgrasses were determined. Salt seretion phenomenon was observedand salt secretion capacity was determinded in the four warm-season turfgrassesunder the salinity stress. Seashore paspalum, the most salt tolerance turfgrass species,and centipedegrass, the salt sensitive species, were selected for the studies ofdifferential photosynthetic responses to salt stress, and the differentially-expressedsalt-responsive proteins in the roots. The main contents and results of this study are asfollows:
1. Salinity stress responses and thresholds in four warm-season turfgrasses
Four warm-season turfgrass species including bermudagrass ‘Tifway’(Cynodondactylon×C. transvaalensis), centipedegrass‘Civil’(Eremochloa ophiuroides),seashore paspalum ‘Salam’(Paspalum vaginatum), and zoysiagrass ‘Matrella’(Zoysia matrella L.Merr) were grown in plastic pots under a series of NaClconcentrations to study the salinity stress responses in turfgrass growth andphysiological parameters. The results indicated that: sashore pspalum has the bestsalinity tolerance. The visual quality, relative growth rate, relative water content,chlorophyll index, Fv/Fm decreased52.2%、75.9%、62.1%、59.4%、72.9%respectively, at36d under600mM of NaCl, and electrolyte leakage increased to 71.5%. The cell membrane damage in seashore paspalum is the lowest whencompared to other three species. Zoysiagrass also showed the good salinity tolerance,with the visual quality, relative growth rate, relative water content, chlorophyll index,Fv/Fm decreased85.2%、68.8%、70.5%、73.9%、49.3%respectively, at36d under600mM of NaCl, and electrolyte leakage increased to81.8%. Bermudagrass andcentipedegrass showed the lowest salinity tolerance, for the treatment of600mMafter36d, both grasses were died. The critical salinity tolerance concentrations foreach parameter were calculated by the regression analysis and Least SignificantDifference (P<0.05). The salinity tolerance thresholds for each turfgrasses are:seashore paspalum261mM, zoysiagrass203mM, bermudagrass135mM andcentipedegrass131mM.
2. Study of salt excretion ability in four warm-season turfgrass
Four warm-season turfgrass species including bermudagrass ‘Tifway’(Cynodondactylon×C. transvaalensis), centipedegrass ‘Civil’(Eremochloa ophiuroides),seashore paspalum ‘Salam’(Paspalum vaginatum), zoysiagrass‘JD-1’(Zoysiajapomca) were grown in plastic pots under300mM NaCl concentration to study theirsalt secretion ability and made the microscopic observation for salt secretion process.The results indicated that: the most salt tolerant seashore paspalum and salt-sensitivecentipedegrass have no function of salt excretion, while zoysiagrass andbermudagrass have the salt excretion function. The salt secretion structures in bothturfgrass are salt glands, and the salt secretion capacity of bermudagrass is higher thanzoysiagrass. At20d of the salt treatment, the salt secretion content of bermudagrass is1.6times higher than zoysiagrass. Mainly due to the salt glands density ofbermudagrass is larger than zoysiagrass. The leaf salt gland number of zoysiagrassaveraged66/mm~2, while the leaf salt gland number of bermudagrass averaged169/mm~2, mermudagrass is2.6times higher than zoysiagrass.
3. Differential photosynthetic responses to salinity stress between seashorepaspalum and centipedegrass in salinity tolerance
This study was to examine photosynthetic responses to salinity stress in twowarm-season turfgrasses differing in salinity tolerance. Salt-tolerant species seashorepaspalum and salt-sensitive species centipedegrass were exposed to salinity at threeNaCl concentrations (0,300and500mM) in a growth chamber. Turf quality, RWC,and Fv/Fm declined while EL increased under the two NaCl regimes for both grassspecies, and the changes were more dramatic in centipedegrass than that in seashorepaspalum, as well as in the higher salinity concentration. Two grass species showeddifferent phytosynthetic responses to salinity stress. The earlier inhibition ofphotosynthesis in seashore paspalum was mainly associated with stomatal closure. Assalinity increased and salinity stress prolonged, the inhibition of photosynthesis inseashore paspalum was mainly associated with non-stomatal factors. The inhibition ofphotosynthesis in centipedegrass was associated with non-stomatal factors at bothsalinity levels. The SDS-PAGE analysis demonstrated the Rubisco large subunit hadno obvious decrease during the whole stress period under the300mM and500mMtreatments in seashore paspalum while significantly decreased in centipedegrass underboth the300mM and500mM treatments. The results indteicated that the superiorsalinity tolerance in seashore paspalum, compared to centipedegrass, could beattributed to its high Pn recovery capacity under salinity stress.
4. Identification of differentially-expressed salt-responsive proteins in roots oftwo perennial grass species contrasting in salinity tolerance
Seashore paspalum and centipedegrass were exposed to salinity stress bywatering the soil with300mM NaCl solution for20d in a growth chamber. Rootswere harvested at20d after salinity treatment. Proteins were extracted and separatedby two-dimensional electrophoresis (2-DE). Root electrolyte leakage (REL) increasedsignificantly while root osmotic potential and root viability decreased significantlyunder salinity treatment for both grass species. However, the extent of change in theabove physiological parameters in seashore paspalum was less pronounced than thosein centipedegrass. Under salinity stress, the abundance of8protein spots significantlyincreased and14significantly decreased in seashore paspalum, while19and16 protein spots exhibited increase and decrease in abundance in centipedegrass,respectively. By comparison of protein spots differentially accumulated between thetwo species,8protein spots that exhibited enhanced abundance in seashore paspalumunder salinity stress were excised from the2-DE gels and subjected to massspectrometry analysis. Seven protein spots were successfully identified. Two of themwere identified as the same protein: ATP synthase Delta chain (ATPS chain);5ofthem were previously confirmed as salt stress-responsive proteins: peroxidase (POD,2.36-fold), cytoplasmic malate dehydrogenase (cMDH,5.84-fold), asorbateperoxidase (APX,4.03-fold) and mitochondrial ATPS chain (2.26-fold and4.78-fold);2of them are novel ones: hypothetical protein LOC100274119(5.01-fold)and Flavoprotein wrbA (2.20-fold). Compared to the8enhanced salt-responsiveprotein spots in seashore paspalum,4corresponding protein spots (cMDH,2ATPSchain, LOC100274119) were expressed and3corresponding protein spots (POD,APX, Flavoprotein wrbA) were not expressed in centipedegrass. The expressedprotein spots in centipedegrass were not significantly changed in their abundance.Immunblotting analysis indicated that POD and ATPS chain were significantlyup-regulated in seashore paspalum at20d of salinity treatment while almost noexpression in both control and salt treatment of centipedegrass. The POD and APXactivities of seashore paspalum were markedly increased while centipedegrass weresignificantly declined at the end of salt treatment. These results indicated that thesuperior salinity tolerance in seashore paspalum, compared to centipedegrass, couldbe associated with its high POD, APX ability and energy metabolism.
1．暖季型草坪草的耐盐阈值
以杂交狗牙根‘Tifway’(Cynodon dactylon×C. transvaalensis)、假俭草‘Civil’(Eremochloa ophiuroides)、海滨雀稗‘Salam’(Paspalum vaginatum)和结缕草‘Matrella’(Zoysia matrella L.Merr)四种暖季型草坪草为材料，研究了不同盐(NaCl)浓度处理对草坪草生长、生理的影响。结果表明：海滨雀稗耐盐性最好，海滨雀稗的目测质量、生长量、叶片相对含水量、叶绿素指数、光化学效率在600mM盐浓度处理36d时分别下降了52.2%、75.9%、62.1%、59.4%、72.9%，相对电导率上升至71.5%，与其他三个草种相比，海滨雀稗各指标仍能维持较高的水平，膜伤害程度相对较小；结缕草耐盐性次之，600mM盐处理36d时目测质量、生长量、叶片相对含水量、叶绿素指数、光化学效率分别下降了85.2%、68.8%、70.5%、73.9%、49.3%，相对电导率上升至81.8%，表现出较好的耐盐性；耐盐性表现较差的为狗牙根和假俭草，在600mM盐浓度处理36d时，两种草的各指标皆下降至0，膜伤害程度达到最大，植物已枯死。以36d时的各生理指标与盐处理浓度建立回归方程，以最小显著差异性检验(LSD0.05)的置信区间分别求出各指标与对照无显著差异的盐浓度临界值。选取最低的临界盐浓度作为该植物的耐盐阈值。求得四个草种的耐盐阈值分别为为：海滨雀稗261mM、结缕草203mM、狗牙根135mM、假俭草131mM。
2．暖季型草坪草的泌盐能力
对杂交狗牙根‘Tifway’(Cynodon dactylon×C. transvaalensis)、假俭草‘Civil’(Eremochloa ophiuroides)、海滨雀稗‘Salam’(Paspalum vaginatum)和上海结缕草‘JD-1’(Zoysia japomca)叶表面盐分测定及活体显微观察发现，最耐盐的海滨雀稗和盐敏感的假俭草没有泌盐功能，而结缕草和狗牙根则具有泌盐功能。通过荧光反射显微镜观察发现，结缕草和狗牙根泌盐结构为盐腺，且狗牙根泌盐量比结缕草要高，在盐处理的20d狗牙根泌盐量为结缕草的1.6倍。主要原因为狗牙根盐腺密度比结缕草大，结缕草叶片盐腺数量平均为66个/mm~2，而狗牙根叶片盐腺数量为169个/mm~2，是结缕草盐腺数量的2.6倍。通过荧光反射显微观察，狗牙根和结缕草的泌盐随着盐处理时间的延长，泌盐量不断增加。但两者盐腺数量在盐胁迫下没有明显变化。狗牙根的泌盐能力高于结缕草，但其抗盐能力却低于结缕草，且耐盐能力最强的海滨雀稗和盐敏感的假俭草中均未发现具有泌盐能力，说明泌盐机制及泌盐能力的大小并不直接决定暖季型草坪草的抗盐能力。
3．海滨雀稗和假俭草对盐胁迫的光合响应
以耐盐的海滨雀稗和盐敏感的假俭草为材料，利用盆栽研究不同浓度盐处理(0,300和500mM NaCl)下的光合生理响应变化。海滨雀稗和假俭草的草坪目测质量、叶片相对含水量及Fv/Fm均随处理时间及处理浓度的递增而呈下降趋势，叶片质膜透性呈上升趋势；盐敏感的假俭草的各指标的变化均明显大于耐盐的海滨雀稗。盐胁迫下两个草种表现出不同的光合响应变化：海滨雀稗在低浓度盐处理初期，光合速率的下降主要是由气孔性因素即气孔关闭引起的胞间CO2浓度降低所造成的，随着处理时间的延长，海滨雀稗光合速率有所恢复，此时海滨雀稗gs有所回升，Ci与对照无显著差异，叶绿素含量、Fv/Fm及Rubisco大亚基均未显著下降。假俭草在两浓度处理下光合速率的下降主要是由非气孔性因素作用的结果，假俭草的光合速率及各生理指标持续下降，Ci持续升高，Rubisco大亚基含量显著下降。海滨雀稗比假俭草抗盐是由于海滨雀稗具有较强的光合恢复能力。
4．海滨雀稗和假俭草根系蛋白表达差异
以抗盐海滨雀稗和盐敏感假俭草为材料，利用2-DE研究两者盐胁迫下根系蛋白差异表达的情况。结果表明，根系渗透势和根系活力在盐胁迫下随着处理时间的延长而不断下降，根系质膜透性随处理时间的延长不断上升。但假俭草各生理指标的变化程度要比海滨雀稗严重。2-DE检测结果表明，海滨雀稗中有8个蛋白点显著上调，14个蛋白点显著降解；假俭草中分别有19和16个蛋白点显著上调和降解。对海滨雀稗8个上调蛋白进行质谱鉴定，7个蛋白点鉴定成功，鉴定结果为：两个蛋白为ATP合成酶链(ATPS链)，五个为盐胁迫响应蛋白，分别是：过氧化物酶(POD，2.36倍)，苹果酸脱氢酶(cMDH，5.84倍)，抗坏血酸过氧化物酶(APX，4.03倍)及线粒体ATP合成酶链(ATPS链，2.26倍和4.78)；2个为新蛋白：hypothetical protein LOC100274119(5.01倍)和FlavoproteinwrbA (2.20倍)。通过对POD和ATP合成酶链进行蛋白免疫印迹验证发现，海滨雀稗在盐处理下，POD和ATPS链的表达明显高于对照，而假俭草对照与处理间无显著差异。本研究结果表明，和假俭草相比，海滨雀稗较高的抗盐性可能与它较高的POD、APX活性及较高的ATP合成能力有关。
Salinity is one of the major abiotic factors limiting plant growth and inducesvarious changes in the growth, physiological and proteomic levels. Understanding themechanisms of salinity resistance and adaptation in turfgrass will provide the crucialreference to select suitable turfgrass variety for salinity soils and imformation forsalinity-tolerant variety development. In this study, existing turfgrass varieties werescreened for the salinity tolerance and the salt tolerance thresholds of fourwarm-season turfgrasses were determined. Salt seretion phenomenon was observedand salt secretion capacity was determinded in the four warm-season turfgrassesunder the salinity stress. Seashore paspalum, the most salt tolerance turfgrass species,and centipedegrass, the salt sensitive species, were selected for the studies ofdifferential photosynthetic responses to salt stress, and the differentially-expressedsalt-responsive proteins in the roots. The main contents and results of this study are asfollows:
1. Salinity stress responses and thresholds in four warm-season turfgrasses
Four warm-season turfgrass species including bermudagrass ‘Tifway’(Cynodondactylon×C. transvaalensis), centipedegrass‘Civil’(Eremochloa ophiuroides),seashore paspalum ‘Salam’(Paspalum vaginatum), and zoysiagrass ‘Matrella’(Zoysia matrella L.Merr) were grown in plastic pots under a series of NaClconcentrations to study the salinity stress responses in turfgrass growth andphysiological parameters. The results indicated that: sashore pspalum has the bestsalinity tolerance. The visual quality, relative growth rate, relative water content,chlorophyll index, Fv/Fm decreased52.2%、75.9%、62.1%、59.4%、72.9%respectively, at36d under600mM of NaCl, and electrolyte leakage increased to 71.5%. The cell membrane damage in seashore paspalum is the lowest whencompared to other three species. Zoysiagrass also showed the good salinity tolerance,with the visual quality, relative growth rate, relative water content, chlorophyll index,Fv/Fm decreased85.2%、68.8%、70.5%、73.9%、49.3%respectively, at36d under600mM of NaCl, and electrolyte leakage increased to81.8%. Bermudagrass andcentipedegrass showed the lowest salinity tolerance, for the treatment of600mMafter36d, both grasses were died. The critical salinity tolerance concentrations foreach parameter were calculated by the regression analysis and Least SignificantDifference (P<0.05). The salinity tolerance thresholds for each turfgrasses are:seashore paspalum261mM, zoysiagrass203mM, bermudagrass135mM andcentipedegrass131mM.
2. Study of salt excretion ability in four warm-season turfgrass
Four warm-season turfgrass species including bermudagrass ‘Tifway’(Cynodondactylon×C. transvaalensis), centipedegrass ‘Civil’(Eremochloa ophiuroides),seashore paspalum ‘Salam’(Paspalum vaginatum), zoysiagrass‘JD-1’(Zoysiajapomca) were grown in plastic pots under300mM NaCl concentration to study theirsalt secretion ability and made the microscopic observation for salt secretion process.The results indicated that: the most salt tolerant seashore paspalum and salt-sensitivecentipedegrass have no function of salt excretion, while zoysiagrass andbermudagrass have the salt excretion function. The salt secretion structures in bothturfgrass are salt glands, and the salt secretion capacity of bermudagrass is higher thanzoysiagrass. At20d of the salt treatment, the salt secretion content of bermudagrass is1.6times higher than zoysiagrass. Mainly due to the salt glands density ofbermudagrass is larger than zoysiagrass. The leaf salt gland number of zoysiagrassaveraged66/mm~2, while the leaf salt gland number of bermudagrass averaged169/mm~2, mermudagrass is2.6times higher than zoysiagrass.
3. Differential photosynthetic responses to salinity stress between seashorepaspalum and centipedegrass in salinity tolerance
This study was to examine photosynthetic responses to salinity stress in twowarm-season turfgrasses differing in salinity tolerance. Salt-tolerant species seashorepaspalum and salt-sensitive species centipedegrass were exposed to salinity at threeNaCl concentrations (0,300and500mM) in a growth chamber. Turf quality, RWC,and Fv/Fm declined while EL increased under the two NaCl regimes for both grassspecies, and the changes were more dramatic in centipedegrass than that in seashorepaspalum, as well as in the higher salinity concentration. Two grass species showeddifferent phytosynthetic responses to salinity stress. The earlier inhibition ofphotosynthesis in seashore paspalum was mainly associated with stomatal closure. Assalinity increased and salinity stress prolonged, the inhibition of photosynthesis inseashore paspalum was mainly associated with non-stomatal factors. The inhibition ofphotosynthesis in centipedegrass was associated with non-stomatal factors at bothsalinity levels. The SDS-PAGE analysis demonstrated the Rubisco large subunit hadno obvious decrease during the whole stress period under the300mM and500mMtreatments in seashore paspalum while significantly decreased in centipedegrass underboth the300mM and500mM treatments. The results indteicated that the superiorsalinity tolerance in seashore paspalum, compared to centipedegrass, could beattributed to its high Pn recovery capacity under salinity stress.
4. Identification of differentially-expressed salt-responsive proteins in roots oftwo perennial grass species contrasting in salinity tolerance
Seashore paspalum and centipedegrass were exposed to salinity stress bywatering the soil with300mM NaCl solution for20d in a growth chamber. Rootswere harvested at20d after salinity treatment. Proteins were extracted and separatedby two-dimensional electrophoresis (2-DE). Root electrolyte leakage (REL) increasedsignificantly while root osmotic potential and root viability decreased significantlyunder salinity treatment for both grass species. However, the extent of change in theabove physiological parameters in seashore paspalum was less pronounced than thosein centipedegrass. Under salinity stress, the abundance of8protein spots significantlyincreased and14significantly decreased in seashore paspalum, while19and16 protein spots exhibited increase and decrease in abundance in centipedegrass,respectively. By comparison of protein spots differentially accumulated between thetwo species,8protein spots that exhibited enhanced abundance in seashore paspalumunder salinity stress were excised from the2-DE gels and subjected to massspectrometry analysis. Seven protein spots were successfully identified. Two of themwere identified as the same protein: ATP synthase Delta chain (ATPS chain);5ofthem were previously confirmed as salt stress-responsive proteins: peroxidase (POD,2.36-fold), cytoplasmic malate dehydrogenase (cMDH,5.84-fold), asorbateperoxidase (APX,4.03-fold) and mitochondrial ATPS chain (2.26-fold and4.78-fold);2of them are novel ones: hypothetical protein LOC100274119(5.01-fold)and Flavoprotein wrbA (2.20-fold). Compared to the8enhanced salt-responsiveprotein spots in seashore paspalum,4corresponding protein spots (cMDH,2ATPSchain, LOC100274119) were expressed and3corresponding protein spots (POD,APX, Flavoprotein wrbA) were not expressed in centipedegrass. The expressedprotein spots in centipedegrass were not significantly changed in their abundance.Immunblotting analysis indicated that POD and ATPS chain were significantlyup-regulated in seashore paspalum at20d of salinity treatment while almost noexpression in both control and salt treatment of centipedegrass. The POD and APXactivities of seashore paspalum were markedly increased while centipedegrass weresignificantly declined at the end of salt treatment. These results indicated that thesuperior salinity tolerance in seashore paspalum, compared to centipedegrass, couldbe associated with its high POD, APX ability and energy metabolism.
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