Investigating the mechanisms underlying phytoprotection by plant growth‐promoting rhizobacteria in Spartina densiflora under metal stress

• Pollution of coasts by toxic metals and metalloids is a worldwide problem for which phytoremediation using halophytes and associated microbiomes is becoming relevant. Metal(loid) excess is a constraint for plant establishment and development, and plant growth promoting rhizobacteria (PGPR) mitigate plant stress under these conditions. However, mechanisms underlying this effect remain elusive. The effect of toxic metal(loid)s on activity and gene expression of ROS‐scavenging enzymes in roots of the halophyte Spartina densiflora grown on real polluted sediments in a greenhouse experiment was investigated.
• Sediments of the metal‐polluted joint estuary of Tinto and Odiel rivers and control, unpollutred samples from the Piedras estuary were collected and submitted to ICP‐OES. Seeds of S. densiflora were collected from the polluted Odiel marshes and grown in polluted and unpolluted sediments. Rhizophere biofilm‐forming bacteria were selected based on metal tolerance and inoculated to S. densiflora and grown for 4 months. Fresh or frozen harvested plants were used for enzyme assays and gene expression studies, respectively.
• Metal excess induced SOD (five‐fold increase), whereas CAT and ascorbate peroxidase displayed minor induction (twofold). A twofold increase of TBARs indicated membrane damage. Our results showed that metal‐resistant PGPR (P. agglomerans RSO6 and RSO7 and B. aryabhattai RSO25) contributed to alleviate metal stress, as deduced from lower levels of all antioxidant enzymes to levels below those of non‐exposed plants. The oxidative stress index (OSI) decreased between 50 and 75% upon inoculation.
• The results also evidenced the important role of PAL, involved in secondary metabolism and/or lignin synthesis, as a pathway for metal stress management in this halophyte upon inoculation with appropriate PGPR, since the different inoculation treatments enhanced PAL expression between 3.75‐ and five‐fold. Our data confirm, at the molecular level, the role of PGPR in alleviating metal stress in S. densiflora and evidence the difficulty of working with halophytes for which little genetic information is available.

     

盐度和淹水对长江口潮滩盐沼植物碳储量的影响

盐生植物是盐沼有机碳储存的"临时库",也是土壤有机碳累积的主要来源,其碳储量大小对盐沼生态系统"碳汇"功能的发挥十分重要。以长江口潮滩本地种芦苇(Phragmites australis)和海三棱藨草(Scirpus mariqueter),及入侵种互花米草(Spartina alterniflora)为研究对象,采用单因素盆栽实验,模拟分析淹水盐度(0、5、10、15、25和35)、淹水深度(0、10、20、40、60cm和80cm)和淹水频率(每天、每3天、每7天、每10天和每15天)变化对各盐生植物地上、地下和总体碳储量大小的影响。研究结果表明,随着淹水盐度增加,芦苇、互花米草和海三棱藨草地上部分与总体碳储量均显著降低。土壤盐度可分别解释其地上部分碳储量变异的47.2%、66.5%和72.7%,与总体碳储量变异的34.7%、45.0%和62.0%。随着淹水深度增加,芦苇地上部分、总体碳储量和海三棱藨草地上部分碳储量均显著降低,其变异的68.6%、28.5%和71.1%可由淹水深度变化(10—80cm)解释。互花米草在80cm淹水深度下仍有较高的地上部分碳储量和总体碳储量。3种盐生植物碳储量对淹水频率变化的响应差异均不显著,所有处理地下部分碳储量差异也未达到显著水平。总体而言,互花米草对水盐胁迫的耐受性要强于本地种芦苇和海三棱藨草。尽管互花米草和芦苇具有相对较高的碳储量,但水盐胁迫对其碳储量的显著抑制作用不容忽视。海三棱藨草碳储量本就不高,输入土壤的有机碳量较为有限,海平面上升及盐水入侵等逆境胁迫会使其对盐沼"碳汇"贡献更加微弱。     

中华补血草内生与根际具ACC脱氨酶活性细菌的筛选及其生物多样性

【目的】获得江苏沿海滩涂盐生药用植物中华补血草内生及根际具有1-氨基环丙烷-1-羧酸(ACC)脱氨酶活性的细菌,研究其遗传多样性和潜在促生活性。【方法】从中华补血草和根际土壤分离筛选具有ACC脱氨酶活性的菌株,对其ACC脱氨酶活性定量检测,通过16S r RNA基因序列分析确定菌株系统发育地位。同时研究其固氮、溶磷、产植物生长素吲哚乙酸(IAA)及耐盐能力。【结果】分离筛选获得18株具有ACC脱氨酶活性的内生与根际细菌,定量检测发现其中有13株菌的ACC脱氨酶含量在20 nmolα-KA/(mg Pr·h)以上,有11株菌可以固氮,7株菌能够解磷,9株菌产生IAA。菌株的Na Cl盐耐受范围多数在0–13%之间。16S r RNA基因测序表明,活性菌株分属于7个属,多样性丰富,节杆菌属(Arthrobacter)为优势类群。其中菌株KLBMP 5180为节杆菌属的潜在新种。【结论】江苏沿海滩涂盐生药用植物中华补血草共生环境中具有丰富多样的具ACC脱氨酶活性的菌株,并存在潜在新物种资源,具有进一步研究价值。     

基于高通量测序分析盐角草根部内生细菌多样性及动态规律

【目的】探讨盐角草根部内生细菌群落多样性特征,揭示内生细菌群落结构在宿主关键发育期动态变化规律。【方法】通过罗氏454高通量测序获得内生细菌16S r RNA片段,然后进行生物信息分析。【结果】共获得20363条16S r RNA基因序列。各样品中可操作分类单元(operational taxonomic units,OTUs)在552–941之间。根部内生细菌群落主要包括4个门,其中Proteobacteri门占主导地位,其余依次是Firmicutes,Actinobacteria,Bacteroidetes。在Proteobacteria门中,Gammaproteobacteria是第一大纲,其后是Betaproteobacteria纲。宿主5个发育时期共同拥有7个细菌属,包括Azomonas,Serratia,Pantoea,Serpens,Pseudomonas,Halomonas,Kushneria。整体上看,Gammaproteobacteria纲在宿主5个发育时期呈现增长趋势。优势菌属在5个发育期存在差异,分别为Delftia,Kushneria,Serratia,Pantoea,Erwinia。所有文库总共含2108个特异OTUs,共同拥有5个相同OTUs。花期OTUs数量最多,结种期内生细菌多样性降低。在宿主的5个发育时期中,土壤p H、月均温和土壤盐含量这3个环境因子组成的集合对其内生细菌群落变化具有显著影响。【结论】盐角草内生细菌群落多样性丰富,宿主发育期决定了内生细菌群落结构。     

Photosynthesis in aquatic adventitious roots of the halophytic stem-succulent Tecticornia pergranulata (formerly Halosarcia pergranulata)

In flood-tolerant species, a common response to inundation is growth of adventitious roots into the water column. The capacity for these roots to become photosynthetically active has received scant attention. The experiments presented here show the aquatic adventitious roots of the flood-tolerant, halophytic stem-succulent, Tecticornia pergranulata (subfamily Salicornioideae, Chenopodiaceae) are photosynthetic and quantify for the first time the photosynthetic capacity of aquatic roots for a terrestrial species. Fluorescence microscopy was used to determine the presence of chloroplasts within cells of aquatic roots. Net O₂ production by excised aquatic roots, when underwater, was measured with varying light and CO₂ regimes; the apparent maximum capacity ( P _max) for underwater net photosynthesis in aquatic roots was 0.45  µ mol O₂ m⁻² s⁻¹. The photosynthetic potential of these roots was supported by the immunolocalization of PsbA, the major protein of photosystem II, and ribulose-1-5-bisphosphate carboxylase/oxygenase (Rubisco) in root protein extracts. Chlorophyllous aquatic roots of T. pergranulata are photosynthetically active, and such activity is a previously unrecognized source of O₂, and potentially carbohydrates, in flooded and submerged plants.     

Effect of temperature, light and salinity on seed germination and radicle growth of the geographically widespread halophyte shrub Halocnemum strobilaceum

BACKGROUND AND AIMS: The small leafy succulent shrub Halocnemum strobilaceum occurs in saline habitats from northern Africa and Mediterranean Europe to western Asia, and it is a dominant species in salt deserts such as those of north-west China. The effects of temperature, light/darkness and NaCl salinity were tested on seed germination, and the effects of salinity were tested on seed germination recovery, radicle growth and radicle elongation recovery, using seeds from north-west China; the results were compared with those previously reported on this species from 'salt steppes' in the Mediterranean region of Spain. METHODS: Seed germination was tested over a range of temperatures in light and in darkness and over a range of salinities at 25 degrees C in the light. Seeds that did not germinate in the NaCl solutions were tested for germination in deionized water. Seeds from which radicles had barely emerged in deionized water were transferred to NaCl solutions for 10 d and then back to deionized water for 10 d to test for radicle growth and recovery. KEY RESULTS: Seeds germinated to higher percentages in light than in darkness and at high than at low temperatures. Germination percentages decreased with an increase in salinity from 0.1 to 0.75 M NaCl. Seeds that did not germinate in NaCl solutions did so after transfer to deionized water. Radicle elongation was increased by low salinity, and then it decreased with an increase in salinity, being completely inhibited by > or = 2.0 M NaCl. Elongation of radicles from salt solutions < 3.0 M resumed after seedlings were transferred to deionized water. CONCLUSIONS: The seed and early seedling growth stages of the life cycle of H. strobilaceum are very salt tolerant, and their physiological responses differ somewhat between the Mediterranean 'salt steppe' of Spain and the inland cold salt desert of north-west China.     

盐土和沙土对新疆常见一年生盐生植物生长和体内矿质组成的影响

选用古尔班通古特沙漠南缘荒漠-绿洲交错带常见的一年生盐生植物盐角草、刺毛碱蓬、叉毛蓬、猪毛菜和碱地肤为材料,比较了它们在原状盐土和沙土中的生长及体内矿质元素组成的差异。结果表明：① 原状盐土0—100 cm各土层的pH值低于沙土,但电导率和含水量明显高于沙土;② 原状盐土中生长的植物干重是沙土中生长植物干重的7—118倍,后者的根冠比是前者的2—6倍。③ 体现肉质化程度的地上部含水率为52%—81%,中低耐盐植物含水率在两种土壤中差异显著,强耐盐植物差异不显著;④ 5种一年生盐生植物地上部氮浓度为11—34 g/kg,在有效氮含量高的盐土上植物氮浓度也高;磷浓度为1—4 g/kg,在有效磷高的盐土上植物磷浓度也较高（盐角草除外）;但沙土中的植物地上部钾浓度明显高于盐土中植株的地上部钾浓度,这与两种土壤在0—60 cm土层中的钾浓度差异相反;⑤ 尽管原状盐土0—100 cm土层中的水溶性钙、镁、钠、氯、硫浓度显著高于沙土,盐土与沙土中生长的植物地上部钠、水溶性氯和硫的浓度比值远远低于土壤中的相应元素浓度的比值,甚至盐土中的植株钙、镁浓度等于或显著低于沙土中生长的植物。表明盐土不仅影响一年生盐生植物的生长,也显著影响这些植物对矿质元素的吸收和累积。一年生盐生植物能够选择性吸收不同生境中的矿质元素。本研究期望为进一步深入研究盐生植物耐盐的适应机制提供依据,也可为植物修复盐碱土的品种选择提供参考。     

Constraints on Salt Marsh Development Following Managed Coastal Realignment: Dispersal Limitation or Environmental Tolerance?

Salt marshes restored through managed coastal realignment (MR) often develop slowly and show persistent differences in vegetation from natural marshes. Development might be constrained by the availability of propagules or poor suitability of the abiotic environment for their establishment. To distinguish between these factors, we compared vegetation colonization and environmental conditions at a salt marsh created by MR at Brancaster, Norfolk, UK, with five reference marshes, varying in age from 30 to circa 6,000 years. After 5 years, plant communities of the MR site remained different from those in mature reference marshes. In contrast, the communities of the youngest reference marsh were not significantly different from mature reference marshes. At the MR site, abundance of perennial and later‐successional species was low and large areas remained unvegetated. These differences are unlikely to be due to dispersal limitation, because 76% of the species from the local species pool colonized the site within 5 years. Although the annuals Salicornia europaea and Suaeda maritima were abundant by year 2, they were not ubiquitous until the end of the study. Tidal elevations of the MR site were suitable for vegetation development, but soil redox potentials were lower than that at the reference sites. Reducing conditions in the MR site appear to be the major cause of vegetation differences from the reference marshes, as they are associated with an abundance of bare ground and a small range of vegetation clusters. Measures to avoid low sediment redox potentials may have a great benefit in some salt marsh restoration projects.     

Control of Na+ and K+ transport in Spergularia marina. III. Relationship between ion uptake and growth at moderate salinity

In this paper, we continue our analysis of Na⁺ and K⁺ uptake by mid-vegetative Spergularia marina (L.) Griseb. plants growing on 0.2x sea water medium, with attention to the relationship of ion uptake and growth. In the first part of the paper, growth analysis techniques are used to compare relative growth rates (RGR) and relative accumulation rates (RAR) for Na⁺ and K⁺. Under constant growth conditions, a high correlation between RGR and RAR indicated that growth and accumulation of both ions were well balanced, resulting in Na⁺ and K⁺ concentrations within the plants which were stable after adjustement to the saline medium. The analysis confirmed the existence of a Na⁺ -related growth stimulation in S. marina and an associated increase in the efficiency of K⁺ utilization for growth. When plants were subjected to more rapid salinization and step changes in the light intensity of the growth chamber, RGR and RAR were again similar, even through the discontinuities in growth conditions, suggesting that growth and ion accumulation were co-regulated rather than simply correlated. The growth analysis data were then transformed to give net uptake rates for Na⁺ and K⁺ and the results were compared to those of isotope studies under similar growth conditions. In roots, the rates estimated by the two techniques differed substantially; net uptake rates reflected primarily growth, while isotope studies indicated a substantial ion exchange rate between mature cells and the growth medium. The rates of transport of either Na⁺ or K⁺ to the shoot were very similar using the two estimation techniques. As the rates measured with isotopes were taken from studies lasting at most a few hours, this suggested a very rapid turnover of the upwardly mobile Na⁺ and K⁺ pools in the roots.