Response Marker Genes Research Articles

Differential expression analysis of a select list of genes in susceptible and resistant heirloom tomatoes with respect to Pseudomonas syringae pv. tomato

Bacterial speck disease of tomato, caused by Pseudomonas syringae pv. tomato (Pst) is one of the most devastating diseases of tomato in the world. To investigate plant responses activated during plant-pathogen interaction, we studied the expression analysis of selected defense-response genes and the microbe or pathogen associated molecular patterns (MAMP or PAMP) triggered immunity (PTI) marker genes in heirloom tomatoes challenged with virulent Pst DC3000. Transcript levels of the defense response genes, including SlPR1a (pathogenesis related proteins), SlPR-Q′b (β-1,3-glucanase), SlGST (Glutathione S-transferase) and peroxidase were up-regulated in the resistant cultivars Orange Strawberry and Amishpaste. The PTI marker genes SlPTI5 (encode a pathogen-inducible ethylene response element-binding protein-like transcription factor) and SlFLS2 (a leucine rich-repeat receptor like kinase) were strongly up-regulated in the incompatible reaction to resistant cultivar Orange Strawberry. On the other hand, transcripts from all tested genes were down-regulated in the compatible interaction in susceptible cultivars Yellow Pear and Brandywine. The induction of defense response and PTI marker genes occurred in the early infection process at 3 days post-inoculation (dpi) and consistent with lower levels of disease severity in resistant cultivars was observed than in susceptible cultivars. Our results contribute to the body of information on bacterial speck disease resistance in an economically important crop tomato.

Dietary deprivation of each essential amino acid induces differential systemic adaptive responses in mice

Dietary deprivation of essential amino acids (EAAs) in mammals is known to cause reductions in food intake and body weight. The aim of this study was to determine whether and how mice respond to deprivation of individual EAA species. Dietary deprivation of any single EAA (not non-EAA) in mice led to progressive weight loss in the order of Ile(-) > Val(-) > Thr(-) > Leu(-) > Trp(-) > His(-) > Phe(-) > Met(-) > Lys(-) , which correlated with the reduction in food intake. Decreased levels of the deprived EAAs as well as increased levels of all or some of the other amino acids were detected in the serum, although these levels differed among the diets examined. Serum biochemistry identified significant increases in creatine phosphokinase, blood urea nitrogen, alanine aminotransferase, and aspartate aminotransferase, and decreases in glucose and triglycerides; computed tomography revealed a marked reduction in abdominal/femoral fat and muscle depots; histology identified diffuse myofiber atrophy in the rectus femoris muscle, all in that approximate order. In contrast, amino acid response, autophagy, and ubiquitination marker genes as well as amino acid transporter genes were induced in both deprived EAA-specific and tissue-specific manners. Dietary deprivation of individual EAAs induced systemic adaptive responses that differed in magnitude and molecular machinery.

The SCF E3 Ligase AtPP2-B11 Plays a Negative Role in Response to Drought Stress in Arabidopsis

Ubiquitination is a eukaryotic post-translational protein modification, implicated in various cellular processes in higher plants, and E3 ligases play an essential role in this cascade. In this study, we identified an Skp1–Cullin–F-box (SCF) E3 ligase encoding gene, Arabidopsis phloem protein 2-B11 (AtPP2-B11; At1g80110), by searching drought-responsive element in the promoter regions of 1,488 putative E3s in the Arabidopsis genome. The expression of AtPP2-B11 in seedlings was remarkably induced with increased duration of drought treatment. Transgenic lines overexpressing AtPP2-B11 exhibited obvious hypersensitivity to drought stress during seed germination and in mature plants. Protein interaction assay with Arabidopsis Skp1 (ASK) proteins showed that AtPP2-B11 strongly interacted with ASK 7, ASK 18, and ASK 19, respectively, indicating that AtPP2-B11 may function as an F-box protein. An AtPP2-B11-interacting protein, AtLEA14, was identified by a yeast two-hybrid screening and bimolecular fluorescence complementation assay. Real-time quantitative RT-PCR and Western blot analysis showed that the transcript and protein levels of AtLEA14 decreased in 35S::AtPP2-B11 transgenic plants subjected to drought conditions. Transgenic Arabidopsis plants overexpressing AtPP2-B11 showed altered expression of abiotic stress response marker genes (COR15a, COR47, ERD10, KIN1, RAB18, and RD22) under both normal and drought conditions. Overall, these results suggested that AtPP2-B11, a SCF E3 ligase, played an important role in response to drought stress as a negative regulator in Arabidopsis.

Prolactin signaling enhances colon cancer stemness by modulating Notch signaling in a Jak2-STAT3/ERK manner

Prolactin (PRL) is a secretory cytokine produced by various tissues. Binding to the cognate PRL receptor (PRLR), it activates intracellular signaling via janus kinase (JAK), extracellular signal-regulated kinase (ERK) and signal transducer and activator of transcription (STAT) proteins. PRL regulates diverse activities under normal and abnormal conditions, including malignancies. Previous clinical data suggest serum PRL levels are elevated in colorectal cancer (CRC) patients. In this study, we first determined the expression of PRL and PRLR in colon cancer tissue and cell lines. Higher levels of PRLR expression were observed in the cancer cells and cell lines compared with normal colonic epithelial cells. Incubation of colon cancer cells with PRL-induced JAK2, STAT3 and ERK1/2 phosphorylation and increased expression of Jagged 1, which is a Notch-1 receptor ligand. Notch signaling regulates CRC stem cell population. We observed increased accumulation of the cleaved/active form of Notch-1 receptor (Notch intracellular domain) and increased expression of Notch responsive genes HEY1, HES1 and stem cell marker genes DCLK1, LGR5, ALDH1 and CD44. Finally, inhibiting PRL induced JAK2-STAT3 and JAK2-ERK1/2 using AG490 and PD98059, respectively, leads to complete abrogation of Notch signaling, suggesting a role for this pathway in regulating CRC stem cells. Together, our results demonstrate that cytokine signaling induced by PRL is active in colorectal cancers and may provide a novel target for therapeutic intervention.

Cloning and functional analysis of Phyllostachys edulis MYB transcription factor PeMYB2

MYB-type transcription factor is one of the largest families in plants, which plays important roles in accepting stress signals from environment and regulating the expression of stress-tolerant genes. In this paper, using homologous cloning and RACE technology, a MYB-type transcription factor, designated PeMYB2, was cloned from Phyllostachys edulis. The results of bioinformatics showed that PeMYB2 is a typical R2R3-MYB. It contained two tandem repeats in its N-terminus, and a membrane protein DUF3651 in its C-terminus. In addition, phylogenetic analysis indicated that PeMYB2 shared the highest homology with 85.98% to OsMYB18 protein from Oryza sativa spp. Japonica. In addition, a yeast one-hybrid assay showed that PeMYB2 could activate the expression of downstream genes. After PeMYB2 was transformed into Arabidopsis thaliana, seven PeMYB2 transgenic Arabidopsis lines were obtained. Phenotypic analysis of the transgenic and wild-type Arabidopsis showed that over-expression of PeMYB2 caused delayed flower or dwarfism in transgenic Arabidopsis. Under the abiotic stress conditions, such as salt and cold stresses, the over-expression of PeMYB2 in Arabidopsis had higher survival rate than the wild-type Arabidopsis. Expression analysis of saline stress response marker genes in the transgenic and wild-type plants under the salt stress condition showed that PeMYB2 regulated the expression of NXH1, SOS1, RD29A, and COR15A. As the result, PeMYB2 might play an important role in various responses to abiotic stresses in P. edulis.

Impact of a functionalized olive oil extract on the uterus and the bone in a model of postmenopausal osteoporosis

The Mediterranean diet rich in fruits, vegetables and olive oil has been related to a lower osteoporosis incidence and accordingly to a reduced fracture risk. These observations might be mediated by the active constituents of extra virgin olive oil, and especially polyphenols. In the context of exploring the features of olive oil active constituents on postmenopausal osteoporosis, an extra virgin olive oil total polyphenolic fraction (TPF) was isolated and its effect on the bone loss attenuation was investigated. Female Lewis rats were ovariectomized and fed a diet enriched with a total phenolic extract of extra virgin olive oil in a concentration of 800mg/kg diet. Oleocanthal, one compound of the polyphenolic fraction, showed a higher relative estrogen receptor binding affinity to the ERα compared to the ERβ. While the TPF only slightly induced the uterine wet weight (490.7±53.7 vs. 432.7±23, p=0.058), TPF regulated estrogen response genes in the uterus (progesterone receptor, antigen identified by monoclonal antibody Ki67, complement C3). Comparing the quantified bone parameters, the oral TPF substitution did not attenuate the ovariectomy-induced bone loss. The administration of extra virgin olive oil polyphenols regulated uterine estrogen response marker genes in an E2-agonistic manner. The bone loss induced by estrogen ablation was not mitigated by treatment with the polyphenolic extract.

GhCAX3 gene, a novel Ca(2+)/H(+) exchanger from cotton, confers regulation of cold response and ABA induced signal transduction.

As a second messenger, Ca2+ plays a major role in cold induced transduction via stimulus-specific increases in [Ca2+]cyt, which is called calcium signature. During this process, CAXs (Ca2+/H+ exchangers) play critical role. For the first time, a putative Ca2+/H+ exchanger GhCAX3 gene from upland cotton (Gossypium hirsutum cv. ‘YZ-1′) was isolated and characterized. It was highly expressed in all tissues of cotton except roots and fibers. This gene may act as a regulator in cotton’s response to abiotic stresses as it could be up-regulated by Ca2+, NaCl, ABA and cold stress. Similar to other CAXs, it was proved that GhCAX3 also had Ca2+ transport activity and the N-terminal regulatory region (NRR) through yeast complementation assay. Over-expression of GhCAX3 in tobacco showed less sensitivity to ABA during seed germination and seedling stages, and the phenotypic difference between wild type (WT) and transgenic plants was more significant when the NRR was truncated. Furthermore, GhCAX3 conferred cold tolerance in yeast as well as in tobacco seedlings based on physiological and molecular studies. However, transgenic plant seeds showed more sensitivity to cold stress compared to WT during seed germination, especially when expressed in N-terminal truncated version. Finally, the extent of sensitivity in transgenic lines was more severe than that in WT line under sodium tungstate treatment (an ABA repressor), indicating that ABA could alleviate cold sensitivity of GhCAX3 seeds, especially in short of its NRR. Meanwhile, we also found that overexpression of GhCAX3 could enhance some cold and ABA responsive marker genes. Taken together, these results suggested that GhCAX3 plays important roles in the cross-talk of ABA and cold signal transduction, and compared to full-length of GhCAX3, the absence of NRR could enhance the tolerance or sensitivity to cold stress, depending on seedling’s developmental stages.

Soil microbiomes vary in their ability to confer drought tolerance to Arabidopsis

Drought is a major constraint on agricultural production. Crop genetic improvement for drought tolerance has received much attention and there is ample information about the ability of specific soil microbes to influence drought tolerance in plants. However, in nature, plants interact simultaneously with an array of beneficial, benign and pathogenic microbes. There is a need to understand the cumulative effect of these multiple interactions on a plant's ability to overcome abiotic stresses such as drought. The objective of this research was to investigate the potential of whole soil microbiomes to help Arabidopsis thaliana plants deal with drought stress under in vivo conditions. A sympatric microbiome (i.e., having a history of exposure to Arabidopsis at a natural site) significantly increased plant biomass under drought conditions, but caused earlier death rates as a consequence of drought; whereas, the two non-sympatric soils did not influence Arabidopsis biomass. Consistent with this, we observed reduced expression levels for several Arabidopsis drought response marker genes (ATDI21, DREB1A, DREB2A, and NCED3) in the sympatric Arabidopsis soil treatment. Pyrosequencing analysis of the three soil microbiomes used in this study identified 84 bacterial OTUs (3% genetic distance) from 41 genera (Burkholderia, Phormidium, Bacillus, Aminobacter, Acidiphilum and among others) that were significantly higher in the sympatric Arabidopsis soil, as compared to the two non-sympatric soils. In conclusion, we have identified a robust set of Arabidopsis-associated microbes that when present in the soil can modify the plant's ability to sense abiotic stress and increase its biomass production.

Abstract 2146: Development of functional assays for p97/VCP inhibition.

Abstract p97 (also called VCP in metazoans and CDC48 in yeast) is a highly conserved, ubiquitously expressed, and essential AAA ATPase. p97 plays a key role in endoplasmic reticulum-associated degradation of misfolded secretory and membrane proteins as well as ubiquitin-dependent turnover of a subset of cytoplasmic substrates of the ubiquitin-proteasome system. p97 also plays a critical role in specific cellular processes including Golgi membrane reassembly, membrane fusion, autophagy and DNA repair mechanisms. p97 expression is elevated in a number of cancer types including prostate cancer, non-small-cell lung carcinoma and leukemia. Given its cellular functions in protein homeostasis and the successful clinical development of proteasome inhibitors, p97 has emerged as an exciting new target for cancer therapy. We used p97 siRNA and a recently identified small molecule p97 inhibitor, N2,N4-dibenzylquinazoline- 2,4-diamine (DBeQ) as tools to develop functional assays suitable for further evaluating the activity of potential p97 inhibitors. A panel of unfolded protein response marker genes that are consistently modulated by p97 siRNA or DBeQ treatment has been identified. In addition, methods to monitor autophagic activity have been developed, including high content analysis of LC3B-GFP puncta formation. Cell viability assays were used to determine the susceptibility of various cancer cell lines to p97 inhibition. These assays will be used to evaluate the effects of potential p97 inhibitors on p97-dependent pathways and anti-proliferative activity. Acknowledgement: Funded by NExT Chemical Biology Consortium, NCI Contract No. HHSN261200800001E. Citation Format: Yihui Shi, Xiaohe Liu, Joy Calaoagan, Steven Samuelsson, Tsui-Fen Chou, Raymond J. Deshaies, Lidia C. Sambucetti. Development of functional assays for p97/VCP inhibition. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2146. doi:10.1158/1538-7445.AM2013-2146

CaWRKY58, encoding a groupI WRKYtranscription factor ofCapsicum annuum, negatively regulates resistance toRalstonia solanacearuminfection

WRKY transcription factors are encoded by large gene families across the plant kingdom. So far, their biological and molecular functions in nonmodel plants, including pepper (Capsicum annuum) and other Solanaceae, remain poorly understood. Here, we report on the functional characterization of a new group I WRKY protein from pepper, termed CaWRKY58. Our data indicate that CaWRKY58 can be localized to the nucleus and can activate the transcription of the reporter β-glucuronidase (GUS) gene driven by the 35S core promoter with two copies of the W-box in its proximal upstream region. In pepper plants infected with the bacterial pathogen Ralstonia solanacearum, CaWRKY58 transcript levels showed a biphasic response, manifested in an early/transient down-regulation and late up-regulation. CaWRKY58 transcripts were suppressed by treatment with methyl jasmonate and abscisic acid. Tobacco plants overexpressing CaWRKY58 did not show any obvious morphological phenotypes, but exhibited disease symptoms of greater severity than did wild-type plants. The enhanced susceptibility of CaWRKY58-overexpressing tobacco plants correlated with the decreased expression of hypersensitive response marker genes, as well as various defence-associated genes. Consistently, CaWRKY58 pepper plants silenced by virus-induced gene silencing (VIGS) displayed enhanced resistance to the highly virulent R. solanacearum strain FJC100301, and this was correlated with enhanced transcripts of defence-related pepper genes. Our results suggest that CaWRKY58 acts as a transcriptional activator of negative regulators in the resistance of pepper to R. solanacearum infection.

Over-expression of a novel JAZ family gene from Glycine soja, increases salt and alkali stress tolerance

Salt and alkali stress are two of the main environmental factors limiting crop production. Recent discoveries show that the JAZ family encodes plant-specific genes involved in jasmonate signaling. However, there is only limited information about this gene family in abiotic stress response, and in wild soybean (Glycine soja), which is a species noted for its tolerance to alkali and salinity. Here, we isolated and characterized a novel JAZ family gene, GsJAZ2, from G. soja. Transcript abundance of GsJAZ2 increased following exposure to salt, alkali, cold and drought. Over-expression of GsJAZ2 in Arabidopsis resulted in enhanced plant tolerance to salt and alkali stress. The expression levels of some alkali stress response and stress-inducible marker genes were significantly higher in the GsJAZ2 overexpression lines as compared to wild-type plants. Subcellular localization studies using a GFP fusion protein showed that GsJAZ2 was localized to the nucleus. These results suggest that the newly isolated wild soybean GsJAZ2 is a positive regulator of plant salt and alkali stress tolerance.

Isolation and functional analysis of <I>GsTIFY11b</I> relevant to salt and alkaline stress from <I>Glycine soja</I>

Using homologous cloning and RT-PCR technology, we isolated a novel TIFY family gene, GsTIFY11b, from Glycine soja L. G07256, a species that is tolerant to saline and alkaline environments. Phylogenetic analysis indicated that GsTIFY11b was closely related to AtTIFY11a with 56% similarity in amino acid identity. Protein sequence analysis showed that GsTIFY11b protein also had conserved TIFY domain, N-terminal domain, and a C-terminal Jas motif. Quantitative realtime PCR analysis indicated that the expression of GsTIFY11b was induced by both saline and alkaline stresses. Two homozygous GsTIFY11b over-expressing transgenic Arabidopsis lines were obtained. Phenotypic analysis of the transgenic and wild-type Arabidopsis indicated that over-expressing GsTIFY11b in Arabidopsis did not enhance plant tolerance to saline and alkaline stresses, whereas it showed an increased sensitivity to saline stress during seed germination and seedling development. Expression analysis of saline stress response marker genes in transgenic and wild-type plants under stress condition indicated that GsTIFY11b regulated the expression of RD29B, KIN1, and DREB. The transient expression of a GsTIFY11b-GFP fusion protein in onion epidermal cells showed that GsTIFY11b was localized to the nucleus, suggesting a role as a transcriptional regulator in the saline stress response pathway.

Expression profiling of putative Eucalyptus grandisdefence marker genes in response to treatment with methyl jasmonate and salicylic acid

Background Eucalyptus species and their hybrids encompass approximately 40% of forestry plantation area in South Africa and contribute significantly to the paper pulp industry due to their favourable wood fibre properties. Eucalypt plantation trees are affected by numerous pathogens during their lifetime, some of which can cause severe losses such as Phytophtora spp and Chrysoporthe spp. Plant defence mechanisms against pathogens is currently better understood in the model plant Arabidopsis thaliana where it has been shown that the salicylic acid (SA) and jasmonic acid (JA) signalling pathways enhance resistance against biotrophic and necrotrophic pathogens, respectively [1] . This process involves the up-regulation of specific defence genes which are considered to be marker (diagnostic) genes for the two signalling pathways [2,3].

Down-Regulation of the Histone Methyltransferase EZH2 Contributes to the Epigenetic Programming of Decidualizing Human Endometrial Stromal Cells

Differentiation of human endometrial stromal cells (HESC) into decidual cells represents a highly coordinated process essential for embryo implantation. We show that decidualizing HESC down-regulate the histone methyltransferase enhancer of Zeste homolog 2 (EZH2), resulting in declining levels of trimethylation of histone 3 on lysine 27 (H3K27me3) at the proximal promoters of key decidual marker genes PRL and IGFBP1. Loss of H3K27me3 was associated with a reciprocal enrichment in acetylation of the same lysine residue, indicating active remodeling from repressive to transcriptionally permissive chromatin. Chromatin immunoprecipitation coupled with DNA microarray analysis demonstrated that decidualization triggers genome-wide changes in H3K27me3 distribution that only partly overlap those observed upon EZH2 knockdown in undifferentiated HESC. Gene ontology revealed that gain of the repressive H3K27me3 mark in response to decidualization and upon EZH2 knockdown in undifferentiated cells was enriched at the promoter regions of genes involved in transcriptional regulation and growth/cell proliferation, respectively. However, loss of the H3K27me3 mark (indicating increased chromatin accessibility) in decidualizing cells and upon EZH2 knockdown occurred at selective loci enriched for genes functionally implicated in responses to stimulus. In agreement, EZH2 knockdown in undifferentiated HESC was sufficient to augment the induction of decidual marker genes in response to cyclic AMP and progesterone signaling. Thus, loss of EZH2-dependent methyltransferase activity in the endometrium is integral to the process of chromatin remodeling that enables the transition from a proliferative to a decidual phenotype in response to differentiation cues.

Similar foliar lesions but opposite hormonal patterns in a tomato mutant impaired in ethylene perception and its near isogenic wild type challenged with ozone

The accumulation of ethylene (ET), hydrogen peroxide (H 2O 2), nitric oxide (NO), salicylic (SA) and jasmonic (JA) acids, as well as the transcriptional activation of JA-biosynthetic and hypersensitive response (HR) marker genes were examined in ET-insensitive Never ripe ( Nr) mutant of tomato ( Lycopersicon esculentum L.) and in its near isogenic wild type (WT, cv. New Yorker) during and after an acute ozone (O 3) fumigation. The scoring of O 3-induced foliar symptoms indicated similar sensitivity between WT and Nr. In both genotypes the O 3-induced ET emission peaked after 3 h of fumigation, although with different intensities between them; H 2O 2 and NO bursts occurred, both showing a biphasic time course. However, SA and JA revealed opposite accumulation patterns: in Nr a high level of SA but a low JA accumulation were observed, whereas in WT a marked evolution of JA but a weak SA accumulation were detected. The transcription of certain JA-biosynthetic and HR-inducible genes was similarly activated by O 3 in the two tomato lines. According to the current models on the molecular events leading to the development of O 3-induced foliar symptoms, here we suggest that the partial ET-insensitivity of Nr would have promoted a SA build-up, leading to cell death and to a reduced JA accumulation. In WT the O 3-induced leaf injury without SA build-up but with JA accumulation would have arisen from the perception of high ET and H 2O 2 levels.

GsTIFY10, a novel positive regulator of plant tolerance to bicarbonate stress and a repressor of jasmonate signaling

Recent discoveries show that TIFY family genes are plant-specific genes involved in the response to several abiotic stresses, also acting as key regulators of jasmonate signaling in Arabidopsis thaliana. However, there is limited information about this gene family in wild soybean, nor is its role in plant bicarbonate stress adaptation completely understood. Here, we isolated and characterized a novel TIFY family gene, GsTIFY10, from Glycine soja. GsTIFY10 could be induced by bicarbonate, salinity stress and the phytohormone JA, both in the leaves and roots of wild soybean. Over-expression of GsTIFY10 in Arabidopsis resulted in enhanced plant tolerance to bicarbonate stress during seed germination, early seedling and adult seedling developmental stages, and the expression levels of some bicarbonate stress response and stress-inducible marker genes were significantly higher in the GsTIFY10 overexpression lines than in wild-type plants. It was also found that GsTIFY10 could repress JA signal transduction. The roots of plants overexpressing GsTIFY10 grew longer than wild-type in the presence of MeJA, and some JA response and JA biosynthesis marker genes were suppressed in the GsTIFY10 overexpression lines. Subcellular localization studies using a GFP fusion protein showed that GsTIFY10 is localized to the nucleus. These results suggest that the newly isolated wild soybean GsTIFY10 is a positive regulator of plant bicarbonate stress tolerance and is also a repressor of jasmonate signaling, from hormone perception to transcriptional activity.

Development of full-length cDNAs from Chinese cabbage (Brassica rapa Subsp. pekinensis) and identification of marker genes for defence response.

Arabidopsis belongs to the Brassicaceae family and plays an important role as a model plant for which researchers have developed fine-tuned genome resources. Genome sequencing projects have been initiated for other members of the Brassicaceae family. Among these projects, research on Chinese cabbage (Brassica rapa subsp. pekinensis) started early because of strong interest in this species. Here, we report the development of a library of Chinese cabbage full-length cDNA clones, the RIKEN BRC B. rapa full-length cDNA (BBRAF) resource, to accelerate research on Brassica species. We sequenced 10 000 BBRAF clones and confirmed 5476 independent clones. Most of these cDNAs showed high homology to Arabidopsis genes, but we also obtained more than 200 cDNA clones that lacked any sequence homology to Arabidopsis genes. We also successfully identified several possible candidate marker genes for plant defence responses from our analysis of the expression of the Brassica counterparts of Arabidopsis marker genes in response to salicylic acid and jasmonic acid. We compared gene expression of these markers in several Chinese cabbage cultivars. Our BBRAF cDNA resource will be publicly available from the RIKEN Bioresource Center and will help researchers to transfer Arabidopsis-related knowledge to Brassica crops.

The HyPRP gene EARLI1 has an auxiliary role for germinability and early seedling development under low temperature and salt stress conditions in Arabidopsis thaliana

The effect of the hybrid proline-rich protein (HyPRP) gene EARLI1 on the rate of germination (germinability) of Arabidopsis seeds and seedling growth under low temperature and salt stress conditions was investigated. EARLI1 was induced during germination in embryonic tissues, and was strongly expressed in certain parts of young seedlings. Comparisons of control, overexpressing (OX), and knockout (KO) lines indicated that higher than wild type levels of EARLI1 improved germinability, root elongation, and reduction of sodium accumulation in leaves under salt stress, as well as germinability under low-temperature stress. Abscisic acid (ABA) contents were relatively low after prolonged salt stress, suggesting that EARLI1 has an ABA-independent effect on germinability under these conditions. Overexpression of EARLI1 during germination enhanced the sensitivity of seeds to exogenously applied ABA, suggesting that EARLI1 has an ABA-dependent negative effect on seed germinability under high ABA stress conditions. Well-known stress response marker genes such as COR15a, KIN1, P5SC1, and RD29 were unaffected whereas P5SC2, RD22, or RAB18 were only slightly affected in OX and KO plants. The pleiotropic effects of EARLI1 during stress and an absence of strong regulatory effects on stress marker genes suggest that this HyPRP gene has an auxiliary role for various stress protection responses in Arabidopsis.

Expression of pathogenesis related genes in response to salicylic acid, methyl jasmonate and 1-aminocyclopropane-1-carboxylic acid in Malus hupehensis (Pamp.) Rehd

BackgroundMany studies have been done to find out the molecular mechanism of systemic acquired resistance (SAR) in plants in the past several decades. Numbers of researches have been carried out in the model plants such as arabidopsis, tobacco, rice and so on, however, with little work done in woody plants especially in fruit trees such as apple. Components of the pathway of SAR seem to be extremely conserved in the variety of species. Malus hupehensis, which is origin in China, is strong resistance with rootstock. In the study, we attempted to make the expression pattern of pathogenesis related (PR) genes which were downstream components of the SAR pathway in response to salicylic acid(SA), methyl jasmonate(MeJA) and 1-aminocyclopropane-1-carboxylic acid(ACC) in Malus hupehensis.FindingsIn order to analyze the expression pattern, the partial sequence of three PR genes from Malus hupehensis, MhPR1, MhPR5 and MhPR8 was isolated. These three PR genes were induced by SA, MeJA and ACC. However, MhPR1, MhPR5 and MhPR8 performed a distinct pattern of expression in different plant organs. MhPR5 and MhPR8 were basal expression in leaves, stems and roots, and MhPR1 was basal expression only in stems. The expression of MhPR1, MhPR5 and MhPR8 was enhanced during the first 48 h post-induced with SA, MeJA and ACC.ConclusionsThe results showed that a distinct pattern of expression of PR genes in Malus hupehensis which differed from the previous reports on model plants arabidopsis, tobacco and rice. MhPR1, MhPR5 and MhPR8 were induced by SA, MeJA and ACC, which were regarded as the marker genes in the SAR response in Malus hupehensis. In contrast with herbal plants, there could be specific signal pathway in response to SA, JA and ET for woody plants.

Gain of deleterious function causes an autoimmune response and Bateson–Dobzhansky–Muller incompatibility in rice

Reproductive isolation plays an important role in speciation as it restricts gene flow and accelerates genetic divergence between formerly interbreeding population. In rice, hybrid breakdown is a common reproductive isolation observed in both intra and inter-specific crosses. It is a type of post-zygotic reproductive isolation in which sterility and weakness are manifested in the F(2) and later generations. In this study, the physiological and molecular basis of hybrid breakdown caused by two recessive genes, hbd2 and hbd3, in a cross between japonica variety, Koshihikari, and indica variety, Habataki, were investigated. Fine mapping of hbd2 resulted in the identification of the causal gene as casein kinase I (CKI1). Further analysis revealed that hbd2-CKI1 allele gains its deleterious function that causes the weakness phenotype by a change of one amino acid. As for the other gene, hbd3 was mapped to the NBS-LRR gene cluster region. It is the most common class of R-gene that triggers the immune signal in response to pathogen attack. Expression analysis of pathogen response marker genes suggested that weakness phenotype in this hybrid breakdown can be attributed to an autoimmune response. So far, this is the first evidence linking autoimmune response to post-zygotic isolation in rice. This finding provides a new insight in understanding the molecular and evolutionary mechanisms establishing post-zygotic isolation in plants.