In our previous reports, an endophytic bacterium, Bacillus cereus KP120 was isolated from the halophyte species Kosteletzkya virginica. In this study, the effect of KP120 colonization on Arabidopsis thaliana seedlings was investigated. Our results showed that inoculation with KP120 could promote the growth of A. thaliana seedlings plants under salt-stress conditions, compared with uninoculated controls. After salt treatment, chlorophyll, proline, the activity of antioxidant enzymes, Indole-3-acetic acid and 1-aminocyclopropane-1-carboxylate-deaminase in plants inoculated were increased significantly but malondialdehyde content was decreased compared with the plants under salt stress lonely. Similarly, under non-salt stress, physiological indices above except for MDA in plants inoculated with KP120 were increased compared with control. B. cereus also induced the up-regulation of key genes involved in IAA biosynthesis, responses, transport, down-regulated expression of genes related with ethylene synthesis and response. Our work principally demonstrates that Bacillus cereus KP120 significantly enhances plant growth and increases plant tolerance to salt stress.

Seashore mallow (Kosteletzkya virginica), as a noninvasive perennial halophytic oilseed-producing dicot, is native from the Gulf to the Atlantic coasts of the U.S. The purpose of our research was to investigate 1-aminocyclopropane-1carboxylic acid deaminase (ACCD) producing endophytic fungi from K.virginica. A total of 59 endophytic fungal strains, isolated from roots in K.virginica of seedlings, were grouped into 12 genera including in Penicillium, Aspergillus, Fusarium, Trichoderma, Rhizopycnis sp., Ceriporia Donk, Trametes sp., Schizophyllum commune sp., Alternaria, Cladosporium, Cylindrocarpon, and Scytalidium according to sequences of ITS. The ACD activity of 10 endophytic fungi isolated was detected. T.asperellum had the highest ACC deaminase activity among all 10 isolated genera of fungal strains, followed by T. viride. Dry weight and fresh weight of plant, plant height, root length, SOD activity, and chlorophyll content of wheat and soybean inoculated with T.asperellum or T. viride was increased compared with non-inoculated control plants under non salt or salt stress. Further analysis showed that T.asperellum or T.viride strains induced downregulation of the expression of ethylene synthesis-related genes such as ACC oxidase (ACO) and ACC synthase (ACS), thereby reducing ethylene synthesis and damage to plants under salt stress. These endophytic fungi can be used as alternative bioinoculants to increase crop yield in saline soil.

Hexokinase (HXK) is a bifunctional enzyme involved in carbohydrate metabolism and sugar signal sensing. HXK gene family has been extensively discussed in many species, while the detailed investigations of the family in Glycine max have yet to be reported. In this study, 17 GmHXK genes (GmHXKs) were identified in the G. max genome and the features of their encoded proteins, conserved domains, gene structures, and cis-acting elements were systematically characterized. The GmHXK2 gene isolated from G. max was firstly constructed into plant expression vector pMDC83 and then transformed with Agrobacterium tumefaciens into Arabidopsis thaliana. The expression of integrated protein was analyzed by Western Blotting. Subcellular localization analysis showed that the GmHXK2 was located on both vacuolar and cell membrane. Under salt stress, seedlings growth was significantly improved in Arabidopsis overexpressing GmHXK2 gene. Furthermore, physiological indicators and expression of salt stress responsive genes involved in K⁺ and Na⁺ homeostasis were significantly lower in GmHXK2-silenced soybean seedlings obtained by virus-induced gene silencing (VIGS) technique under salt stress compared with the control plants. Our study showed that GmHXK2 gene played an important role in resisting salt stress, which suggested potential value for the genetic improvement of abiotic resistant crops.

The CHX (cation/H⁺ exchanger) family plays an important role in the transmembrane transport of cation/H⁺ in plants. The aim of this study was to identify and functionally analyze the KvCHX gene in the halophyte Kosteletzkya virginica to investigate its role in regulating the K⁺/Na⁺ ratio under salinity tolerance. Based on a partial gene sequence of EST from K. virginica, the full-length DNA sequence of the KvCHX gene was obtained using genome walking technology. Structural analysis and phylogenetic relationship analysis showed that the KvCHX gene was closely related to the AtCHX17 gene. The KvCHX overexpression vector was successfully constructed and transformed into Arabidopsis via floral dipping. Arabidopsis seedlings overexpressing KvCHX showed an enhanced tolerance to salt stress compared with wild-type plants. Transgenic Arabidopsis seedlings grew better under K⁺ deficiency than WT. The results showed that KvCHX could promote the uptake of K⁺, increase the ratio of K⁺/Na⁺, and promote the growth of plants under K⁺ deficiency and treatment with NaCl solution. KvCHX is involved in K⁺ transport and improves plant salt tolerance by coordinating K⁺ acquisition and homeostasis.

Cadmium (Cd) is one of the most toxic heavy metal pollutants known to accumulate in the soil environment and pose a threat to public health. The tolerance of endophytic fungi isolated from Kosteletzkya virginica to cadmium was determined by the Oxford cup method. Trichoderma asperellum displayed the highest tolerance to cadmium on potato dextrose agar (PDA) medium and was selected subsequently for inoculating maize seedlings. Green fluorescein protein (GFP) labeled T. asperellum colonised in the roots of maize seedlings. Under Cd stress, the plant height, dry weight, fresh weight, root length, chlorophyll content and antioxidant enzyme activity of maize seedlings inoculated with T. asperellum were significantly higher than those of the control. T. asperellum also contributed to the up-regulated expression of some key genes in roots and leaves of maize seedlings, involving nitrogen absorption and metabolism to promote maize growth and Cd detoxification, accumulation, distribution, etc. for Cd transport to increase resistance to Cd.

Previous studies show that 1-aminocyclopropane-1-carboxylate (ACC) deaminase can facilitate the growth and stress tolerance of hosts by reducing ethylene levels. In this study, the acdS gene encoding ACC deaminase from Bacillus cereus (HK012) was cloned and transformed into tobacco (Nicotiana tabacum L.) by the leaf disc method using Agrobacterium. Molecular detection and physiological analysis of the transgenic tobacco plants were performed. Our results showed the acdS gene was integrated into the tobacco genome and fluorescence microscopy showed that the fusion protein was located on the cell membrane of tobacco root. Compared with control, the transgenic plants showed increases in plant height, root length, dry weight, fresh weight and chlorophyll content; and significant increases in the concentration of proline of 55.15% and 42.7% under salt stress conditions (150 mmol L⁻¹ and 300 mmol L⁻¹ NaCl, respectively). The superoxide dismutase, peroxidase, catalase and ACC deaminase activities of transgenic tobacco were higher than those of control tobacco at 150 and 300 mmol L⁻¹ salt concentrations. Transgenic tobacco seedlings expressing the acdS gene of B. cereus HK012 showed higher salt tolerance than the control plants. The obtained results suggest that the acdS gene of B. cereus can be used to promote salt tolerance in glycophytes by using biotechnology strategies.