BcMF11, a novel non-coding RNA gene from Brassica campestris, is required for pollen development and male fertility

Key message

BcMF11 as a non-coding RNA gene has an essential role in pollen development, and might be useful for regulating the pollen fertility of crops by antisense RNA technology.

Abstract

We previously identified a 828-bp full-length cDNA of BcMF11, a novel pollen-specific non-coding mRNA-like gene from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). However, little information is known about the function of BcMF11 in pollen development. To investigate its exact biological roles in pollen development, the BcMF11 cDNA was antisense inhibited in transgenic Chinese cabbage under the control of a tapetum-specific promoter BcA9 and a constitutive promoter CaMV 35S. Antisense RNA transgenic plants displayed decreasing expression of BcMF11 and showed distinct morphological defects. Pollen germination test in vitro and in vivo of the transgenic plants suggested that inhibition of BcMF11 decreased pollen germination efficiency and delayed the pollen tubes’ extension in the style. Under scanning electron microscopy, many shrunken and collapsed pollen grains were detected in the antisense BcMF11 transgenic Chinese cabbage. Further cytological observation revealed abnormal pollen development process in transgenic plants, including delayed degradation of tapetum, asynchronous separation of microspore, and aborted development of pollen grain. These results suggest that BcMF11, as a non-coding RNA, plays an essential role in pollen development and male fertility.     

Expression Profiling of the <Emphasis Type="Italic">CSDP</Emphasis> Transcription Factor Gene Family Points to Roles in Organ Development and Abiotic Stress Response in Tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis> L.)

The cold shock domain proteins (CSDPs) are small group of nucleic acid-binding proteins that act as RNA chaperones in growth regulation, development, and stress adaptation in plants. The functions of CSDPs have been studied in Arabidopsis (Arabidopsis thaliana), rice (Oryza sativa), wheat (Triticum aestivum), and Chinese cabbage (Brassica rapa). To gain insight into the function of CSDPs in tomato (Solanum lycopersicum), we performed a genome-wide analysis of CSDPs through in silico characterization and expression profiling in different organs and in response to different abiotic stress and phytohormone treatments. We identified five non-redundant SlCSDP genes. The evolutionary analysis and phylogenetic classification indicated that tomato CSDPs are more closely related to potato than those of others. The five SlCSDP genes are distributed on four of the 12 tomato chromosomes and no segmental or tandem duplication events are detected among them. Expression analysis showed broad expression patterns with strong expression in fruit development and ripening. Expression of individual SlCSDP genes was significantly altered by stress and phytohormone treatments. SlCSDP2, SlCSDP3, and SlCSDP4 were highly induced by all four abiotic stresses and by phytohormone treatment in tomato. These findings provide a foundation for future research towards functional biological roles of CSDP gene in particular to develop tomato cultivars with large size, early ripening, and abiotic stress tolerance.     

AtHSP17.8 overexpression in transgenic lettuce gives rise to dehydration and salt stress resistance phenotypes through modulation of ABA-mediated signaling

Key message

Transgenic Arabidopsis and lettuce plants overexpressing AtHSP17.8 showed ABA-hypersensitive but abiotic stress-resistant phenotypes. ABA treatment caused a dramatic induction of early ABA-responsive genes in AtHSP17.8 -overexpressing transgenic lettuce.

Abstract

Plant small heat shock proteins function as chaperones in protein folding. In addition, they are involved in responses to various abiotic stresses, such as dehydration, heat and high salinity in Arabidopsis. However, it remains elusive how they play a role in the abiotic stress responses at the molecular level. In this study, we provide evidence that Arabidopsis HSP17.8 (AtHSP17.8) positively regulates the abiotic stress responses by modulating abscisic acid (ABA) signaling in Arabidopsis, and also in lettuce, a heterologous plant when ectopically expressed. Overexpression of AtHSP17.8 in both Arabidopsis and lettuce leads to hypersensitivity to ABA and enhanced resistance to dehydration and high salinity stresses. Moreover, early ABA-responsive genes, ABI1, ABI5, NCED3, SNF4 and AREB2, were rapidly induced in AtHSP17.8-overexpressing transgenic Arabidopsis and lettuce. Based on these data, we propose that AtHSP17.8 plays a crucial role in abiotic stress responses by positively modulating ABA-mediated signaling in both Arabidopsis and lettuce. Moreover, our results suggest that stress-tolerant lettuce can be engineered using the genetic and molecular resources of Arabidopsis.     

Phosphate-solubilizing soil yeast Meyerozyma guilliermondii CC1 improves maize (Zea mays L.) productivity and minimizes requisite chemical fertilization

Aim

Phosphate-solubilizing yeasts have been under-exploited in eco-friendly maize cultivation. In this regard, soil yeasts Meyerozyma guilliermondii CC1, Rhodotorula mucilaginosa CC2 and M. caribbica CC3 were investigated for their plant growth-promoting (PGP) activities.

Methods

Soil yeasts were isolated and characterized. Maize (Zea mays L. cv. Tainong No.1) and Chinese cabbage (Brassica rapa L. cv. Pekinensis) were used for seed bioassay. Growth-promoting effects of yeasts under greenhouse conditions were evaluated using maize and lettuce (Lactuca sativa L. cvs. Capitata and Taiwan sword leaf). Ultimately, M. guilliermondii CC1 was tested on field-grown maize; treatments included full-dose chemical fertilizers (CF), yeast (CC1), half-dose chemical fertilizers (½CF), CC1?+?½CF and control. Nutrient uptake, growth, and yield of maize and rhizospheric soil microbes were estimated.

Results

Strain M. guilliermondii CC1 exhibited better seed vigor index in maize and Chinese cabbage. CC1?+?½CF significantly improved the dry-weights, and nutrient uptakes of maize and sword leaf lettuce under greenhouse conditions. In field, CC1?+?½CF application exerted a pronounced effect on growth of maize, cob yield, nutrient-uptake and rhizospheric soil microbial counts.

Conclusion

Our results validated superior biochemical potency and PGP traits of M. guilliermondii CC1 that reduced requisite chemical fertilizer application without affecting the optimal productivity in maize.     

A Chinese cabbage (Brassica campetris subsp. Chinensis) τ-type glutathione-S-transferase stimulates Arabidopsis development and primes against abiotic and biotic stress

The beneficial root-colonizing fungus Piriformospora indica stimulates root development of Chinese cabbage (Brassica campestris subsp. Chinensis) and this is accompanied by the up-regulation of a τ-class glutathione (GSH)-S-transferase gene (BcGSTU) (Lee et al. 2011) in the roots. BcGSTU expression is further promoted by osmotic (salt and PEG) and heat stress. Ectopic expression of BcGSTU in Arabidopsis under the control of the 35S promoter results in the promotion of root and shoot growth as well as better performance of the plants under abiotic (150 mM NaCl, PEG, 42?°C) and biotic (Alternaria brassicae infection) stresses. Higher levels of glutathione, auxin and stress-related (salicylic and jasmonic acid) phytohormones as well as changes in the gene expression profile result in better performance of the BcGSTU expressors upon exposure to stress. Simultaneously the plants are primed against upcoming stresses. We propose that BcGSTU is a target of P. indica in Chinese cabbage roots because the enzyme participates in balancing growth and stress responses, depending on the equilibrium of the symbiotic interaction. A comparable function of BcGST in transgenic Arabidopsis makes the enzyme a valuable tool for agricultural applications.     

The <Emphasis Type="Italic">Populus trichocarpa PtHSP17.8</Emphasis> involved in heat and salt stress tolerances

Key message

PtHSP17.8 was regulated by various abiotic stresses. Overexpression of PtHSP17.8 enhanced the tolerance to heat and salt stresses through maintain ROS homeostasis and cooperate with stress-related genes in Arabidopsis.

Abstract

Small heat shock proteins (sHSPs) play important roles in response to diverse biotic and abiotic stresses, especially in heat tolerance. However, limited information is available on the stress tolerance roles of sHSPs in woody species. To explore the function of sHSPs in poplar, we isolated and characterized PtHSP17.8 from Populus trichocarpa. Phylogenetic analysis and subcellular localization revealed that PtHSP17.8 was a cytosolic class I sHSP. The gene expression profile of PtHSP17.8 in various tissues showed that it was significantly accumulated in stem and root, which was consistent with the GUS expression pattern driven by promoter of PtHSP17.8. The expression of PtHSP17.8 could be induced by various abiotic stresses and significantly activated by heat stress. Overexpression of PtHSP17.8 enhanced the tolerance to heat and salt stresses in Arabidopsis. The seedling survival rate, root length, relative water content, antioxidative enzyme activities, proline, and soluble sugar content were increased in transgenic Arabidopsis under heat and salt stresses, but not in normal condition. The co-expression network of PtHSP17.8 were constructed and demonstrated many stress responsive genes included. The stress-related genes in the co-expression network were up-regulated in the PtHSP17.8 overexpression seedlings. These results suggest that PtHSP17.8 confers heat and salt tolerances in plants.

     

Expression of hsp70, hsp100 and ubiquitin in Aloe barbadensis Miller under direct heat stress and under temperature acclimation conditions

Key message

The study determined the tolerance of Aloe vera to high temperature, focusing on the expression of hsp70 , hsp100 and ubiquitin genes. These were highly expressed in plants acclimated at 35 °C prior to a heat shock of 45 °C.

Abstract

Aloe barbadensis Miller (Aloe vera), a CAM plant, was introduced into Chile in the semiarid IV and III Regions, which has summer diurnal temperature fluctuations of 25 to 40 °C and annual precipitation of 40 mm (dry years) to 170 mm (rainy years). The aim of this study was to investigate how Aloe vera responds to water and heat stress, focusing on the expression of heat shock genes (hsp70, hsp100) and ubiquitin, which not studied before in Aloe vera. The LT₅₀ of Aloe vera was determined as 53.2 °C. To study gene expression by semi-quantitative RT-PCR, primers were designed against conserved regions of these genes. Sequencing the cDNA fragments for hsp70 and ubiquitin showed a high identity, over 95 %, with the genes from cereals. The protein sequence of hsp70 deduced from the sequence of the cDNA encloses partial domains for binding ATP and the substrate. The protein sequence of ubiquitin deduced from the cDNA encloses a domain for interaction with the enzymes E2, UCH and CUE. The expression increased with temperature and water deficit. Hsp70 expression at 40–45 °C increased 50 % over the controls, while the expression increased by 150 % over the controls under a water deficit of 50 % FC. The expression of all three genes was also studied under 2 h of acclimation at 35 or 40 °C prior to a heat shock at 45 °C. Under these conditions, the plants showed greater expression of all genes than when they were subjected to direct heat stress.     

Plant growth promoting rhizobacterium Proteus vulgaris JBLS202 stimulates the seedling growth of Chinese cabbage through indole emission

Backgrounds and aims

The functional role of volatile indole in interaction between rhizobacteria and plant remains unknown. In this study, we investigated the functional role of the volatile indole emitted by rhizobacterial strain Proteus vulgaris JBLS202 in plant growth promoting activity.

Methods

P. vulgaris strain JBLS202 was used to study the role of volatile organic compounds (VOCs) on growth stimulation of Chinese cabbage (cabbage) at seedling stage. SPME-GC/MS analysis employed to identify headspace VOCs emitted from the rhizobacterium. Synthetic indole was assayed at various concentrations for the growth stimulation of cabbage and the emission of indole from the bacterized cabbage seeds was identified.

Results

P. vulgaris JBLS202 promoted the growth of cabbage via volatiles in a dose-dependent manner. VOC emission assay by SPME-GC/MS revealed that indole was a major headspace volatile compound emitted from the rhizobacterium. Moreover, the growth of cabbage was promoted significantly in the presence of 0.63 μg of synthetic indole. The vigor index and fresh weight of the seedlings were increased by 39.9 % and 32.6 %, respectively when the seeds of cabbage were bacterized with P. vulgaris JBLS202 cells (1?×?10⁷?CFU/ml). The emission of indole from the bacterized seeds was demonstrated by SPME-GC/MS.

Conclusions

Results indicated that either synthetic or biological/bacterial indole could increase the growth of cabbage significantly. Though the molecular biological role of indole in plant growth promotion remains to be investigated, this is the first report on detailed interaction between bacterial indole and plants.     

Overexpression of a novel salt stress-induced glycine-rich protein gene from alfalfa causes salt and ABA sensitivity in Arabidopsis

Key message

We cloned a novel salt stress-induced glycine-rich protein gene ( MsGRP ) from alfalfa. Its overexpression retards seed germination and seedling growth of transgenic Arabidopsis after salt and ABA treatments.

Abstract

Since soil salinity is one of the most significant abiotic stresses, salt tolerance is required to overcome salinity-induced reductions in crop productivity. Many glycine-rich proteins (GRPs) have been implicated in plant responses to environmental stresses, but the function and importance of some GRPs in stress responses remain largely unknown. Here, we report on a novel salt stress-induced GRP gene (MsGRP) that we isolated from alfalfa. Compared with some glycine-rich RNA-binding proteins, MsGRP contains no RNA recognition motifs and localizes in the cell membrane or cell wall according to the subcellular localization result. MsGRP mRNA is induced by salt, abscisic acid (ABA), and drought stresses in alfalfa seedlings, and its overexpression driven by a constitutive cauliflower mosaic virus-35S promoter in Arabidopsis plants confers salinity and ABA sensitivity compared with WT plants. MsGRP retards seed germination and seedling growth of transgenic Arabidopsis plants after salt and ABA treatments, which implies that MsGRP may affect germination and growth through an ABA-dependent regulation pathway. These results provide indirect evidence that MsGRP plays important roles in seed germination and seedling growth of alfalfa under some abiotic stress conditions.