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Symbiotic nitrogen fixation (SNF) in legume nodules is a highly energy demanding process, fuelled by plant-supplied carbohydrates mainly in the form of sucrose. In this study, we have combined molecular and biochemical approaches in order to study the spatial and temporal organisation of sucrose metabolism in nitrogen-fixing nodules of the model legume Lotus japonicus, with an emphasis on the neglected role of alkaline/neutral invertase. For this purpose, a full-length cDNA clone coding for an alkaline/neutral invertase isoform, termed LjInv1, was identified in a L. japonicus mature nodule cDNA libraries. Alkaline/neutral invertase activity was also found to be the predominant invertase activity in mature nodules. Real-time reverse-transcription polymerase chain reaction analysis was used in order to study the temporal expression patterns of LjInv1 in parallel with genes encoding acid invertase and sucrose synthase (SuSy) isoforms, and enzymes involved in the subsequent hexose partitioning including hexokinase, phosphoglucomutase (PGM) and phosphoglucose isomerase (PGI). The spatial organisation of sucrose metabolism was studied by in situ localisation of LjInv1 transcripts and alkaline/neutral invertase activity, and SuSy protein during nodule development. Furthermore, the spatial organisation of hexose metabolism was investigated by histochemical localisation of hexokinase, PGM and PGI activities in mature nodules. The results considered together indicate that alkaline/neutral invertase could contribute to both the Glc-1-P and Glc-6-P pools in nodules, fuelling both biosynthetic processes and SNF. Furthermore, transcript profiling analysis revealed that genes coding for hexokinase and putative plastidic PGM and PGI isoforms are upregulated during the early stages of nodule development, while the levels of transcripts corresponding to cytosolic PGM and PGI isoforms remained similar to uninfected roots, indicating a possible role of LjInv1 in producing hexoses for starch production and other biosynthetic processes in developing nodules.
Plant Mol Biol 2006 Sep
PMID:Spatial and temporal organization of sucrose metabolism in Lotus japonicus nitrogen-fixing nodules suggests a role for the elusive alkaline/neutral invertase. 1689 73

Rice production is known to be severely affected by virus transmitting rice pests, brown planthopper (BPH) and green leafhopper (GLH) of the order hemiptera, feeding by phloem abstraction. ASAL, a novel lectin from leaves of garlic (Allium sativum) was previously demonstrated to be toxic towards hemipteran pests when administered in artificial diet as well as in ASAL expressing transgenic plants. In this report ASAL was targeted under the control of phloem-specific Agrobacterium rolC and rice sucrose synthase-1 (RSs1) promoters at the insect feeding site into popular rice cultivar, susceptible to hemipteran pests. PCR, Southern blot and C-PRINS analyses of transgenic plants have confirmed stable T-DNA integration and the transgenes were co-segregated among self-fertilized progenies. The T(0) and T(1) plants, harbouring single copy of intact T-DNA expression cassette, exhibit stable expression of ASAL in northern and western blot analyses. ELISA showed that the level of expressed ASAL was as high as 1.01% of total soluble protein. Immunohistofluorescence localization of ASAL depicted the expected expression patterns regulated by each promoter type. In-planta bioassay studies revealed that transgenic ASAL adversely affect survival, growth and population of BPH and GLH. GLH resistant T(1) plants were further evaluated for the incidence of tungro disease, caused by co-infection of GLH vectored Rice tungro bacilliform virus (RTBV) and Rice tungro spherical virus (RTSV), which appeared to be dramatically reduced. The result presented here is the first report of such GLH mediated resistance to infection by RTBV/RTSV in ASAL expressing transgenic rice plant.
Plant Mol Biol 2006 Nov
PMID:A novel approach for developing resistance in rice against phloem limited viruses by antagonizing the phloem feeding hemipteran vectors. 1694 Dec 13

Nitrogen fixation (NF) in legume nodules is very sensitive to environmental constraints. Nodule sucrose synthase (SS; EC 2.4.1.13) has been suggested to play a crucial role in those circumstances because its downregulation leads to an impaired glycolytic carbon flux and, therefore, a depletion of carbon substrates for bacteroids. In the present study, the likelihood of SS being regulated by oxidative signaling has been addressed by the in vivo supply of paraquat (PQ) to nodulated pea plants and the in vitro effects of oxidizing and reducing agents on nodule SS. PQ produced cellular redox imbalance leading to an inhibition of NF. This was preceded by the downregulation of SS gene expression, protein content, and activity. In vitro, oxidizing agents were able to inhibit SS activity and this inhibition was completely reversed by the addition of dithiothreitol. The overall results are consistent with a regulation model of nodule SS exerted by the cellular redox state at both the transcriptional and post-translational levels. The importance of such mechanisms for the regulation of NF in response to environmental stresses are discussed.
Mol Plant Microbe Interact 2008 May
PMID:Evidence for transcriptional and post-translational regulation of sucrose synthase in pea nodules by the cellular redox state. 1839 22

The first step in sucrose use by maize kernels produces fructose, regardless of whether the initial reaction is catalyzed by an invertase or the reversible sucrose synthase. This fructose can enter subsequent metabolism via hexokinase, or in maize kernels, by a sorbitol dehydrogenase that reversibly converts fructose + NADH to sorbitol + NAD. High levels of SDH activity suggest that kernels synthesize considerable amounts of sorbitol, but the molecular mechanism and functional role for this process have remained equivocal. To gain insights on the role of sorbitol synthesis in maize endosperm we cloned and characterized the transcriptional control of the maize sorbitol dehydrogenase (Sdh1) gene. Data indicated that Sdh1 was essentially kernel- and endosperm-specific, with maximal expression at both the mRNA and enzyme activity levels during early kernel development. Expression was elevated in high-sugar mutants (sugary1, shrunken2), also by sugar injections, and was more pronounced when transfected tissues were incubated at low oxygen concentrations. Control of Sdh1 expression in our transient assays was largely dependent on the first intron of Sdh1. We speculate that SDH activity may represent an adaptation to the high-sugar/low-oxygen environment of the endosperm. Under these conditions, the NADH-dependent reduction of fructose to sorbitol would regenerate NAD[+], thus contributing to the maintenance of the redox and energy status of the cell.
Plant Mol Biol 2008 Oct
PMID:Sugar levels modulate sorbitol dehydrogenase expression in maize. 1856 93

The regulation of seed oil synthesis in rapeseed is largely unknown. In this study, Arabidopsis microarray was used to analyze the gene differential expression of the immature seeds 30 days after flowering of a high oil Brassica napus, H105, whose oil content was 46.04 +/- 1.42, 53.94 +/- 1.35 and 53.09 +/- 1.35% when planted in Nanjing (altitude: 8.9 m), Xining (altitude: 2261.2 m) and Lhasa (altitude: 3658 m), respectively. Transcript levels of 363 genes and 421 genes were altered twofold or more for H105 planted in Xining and Lhasa compared to that in Nanjing, respectively. Together, there were 53 common up-regulated and 42 common down-regulated expression transcripts shared by H105 planted in Xining and Lhasa compared to that in Nanjing. Some important genes, such as sucrose synthase, pyruvate kinase and 6-phosphogluconate dehydrogenase which related to sugar metabolism were identified common up-regulated in higher oil content H105. These results revealed the expressional disciplinarian of correlative genes, and provided important information of the molecular genetic mechanism of oil content difference of rapeseed. In addition, these differential expression genes could be suitable as targets for genetic improvement of seed oil content.
Mol Biol Rep 2009 Nov
PMID:Microarray analysis of gene expression in seeds of Brassica napus planted in Nanjing (altitude: 8.9 m), Xining (altitude: 2261.2 m) and Lhasa (altitude: 3658 m) with different oil content. 1921 39

A full-length cDNA clone encoding sucrose synthase (SS; EC 2.4.1.13) was isolated from muskmelon (Cucumis melo L.) by RT-PCR and RACE. The clone, designated as CmSS1, contains 2,585 nucleotides with an open reading frame of 2,412 nucleotides. The deduced 804 amino acid sequence showed high identities with other plant sucrose synthase. Real time PCR analysis indicated that CmSS1 expression differed among root, stem, leaf, flower and fruit tissues. The analysis during fruit development indicated that CmSS1 mRNA showed its maximum level at 5 days after pollination (DAP) and decreased gradually during fruit development until its minimum level in mature fruit. The sucrose content was very low in fruit before 20 DAP but increased dramatically between 20 and 30 DAP during fruit development. However, SS activities in both direction of sucrose synthesis and sucrose cleavage were very low and changed little during fruit development, suggesting that SS may play little role in determining sucrose accumulation during muskmelon fruit.
Mol Biol Rep 2010 Feb
PMID:Cloning and characterization of a sucrose synthase-encoding gene from muskmelon. 1941 24

An alfalfa cDNA library induced by salt stress was constructed by suppression subtraction hybridization (SSH) technology. Total RNA from 10-day-old seedlings was used as a "driver," and total RNA from seedlings induced by salt was used as a "tester". One hundred and nineteen clones identified as positive clones by reverse Northern dot-blotting resulted in 82 uni-ESTs comprised of 16 contigs and 66 singletons. Blast analysis of deduced protein sequences revealed that 51 ESTs had identity similar to proteins with known function, while 24 could not be annotated at all. Most of the annotated sequences were homologous to genes involved in abiotic or biotic stress in plants. Among these proteins, beta-amylase, fructose-1,6-bisphosphate, aldolase, and sucrose synthase are related to osmolyte synthesis; a CCCH-type zinc finger protein, DNA binding protein, His-Asp phosphotransfer protein, and the RelA/SpoT protein partake in transcription regulation and signal transduction; and ribulose-l,5-bisphosphate carboxylase/oxygenase, chlorophyll a/b binding proteins, and an early light-inducible proteins are related to photosynthesis. In addition, several ESTs, similar to genes from other plant species, closely involved in salt stress were isolated from alfalfa, such as an aquaporin protein, a late embryogenesis-abundant protein, and glutathione peroxidase.
Mol Biol Rep 2010 Feb
PMID:Screening of genes induced by salt stress from Alfalfa. 1957 13

Regulation of symbiotic nitrogen fixation (SNF) during drought stress is complex and not yet fully understood. In the present work, the involvement of nodule C and N metabolism in the regulation of SNF in Medicago truncatula under drought and a subsequent rewatering treatment was analyzed using a combination of metabolomic and proteomic approaches. Drought induced a reduction of SNF rates and major changes in the metabolic profile of nodules, mostly an accumulation of amino acids (Pro, His, and Trp) and carbohydrates (sucrose, galactinol, raffinose, and trehalose). This accumulation was coincidental with a decline in the levels of bacteroid proteins involved in SNF and C metabolism, along with a partial reduction of the levels of plant sucrose synthase 1 (SuSy1). In contrast, the variations in enzymes related to N assimilation were found not to correlate with the reduction in SNF, suggesting that these enzymes do not have a role in the regulation of SNF. Unlike the situation in other legumes such as pea and soybean, the drought-induced inhibition of SNF in M. truncatula appears to be caused by impairment of bacteroid metabolism and N(2)-fixing capacity rather than a limitation of respiratory substrate.
Mol Plant Microbe Interact 2009 Dec
PMID:Carbon metabolism and bacteroid functioning are involved in the regulation of nitrogen fixation in Medicago truncatula under drought and recovery. 1988 22

Annexins belong to a multigene family of Ca(2+) dependent, phospholipid and cytoskeleton binding proteins. They have been shown to be upregulated under various stress conditions. We generated transgenic cotton plants expressing mustard annexin (AnnBj1), which showed enhanced tolerance towards different abiotic stress treatments like sodium chloride, mannitol, polyethylene glycol and hydrogen peroxide. The tolerance to these treatments was associated with decreased hydrogen peroxide levels and enhanced total peroxidase activity, enhanced content of osmoprotectants- proline and sucrose in transgenic plants. They showed higher retention of total chlorophyll and reduced TBARS in leaf disc assays with stress treatments, and decreased hydrogen peroxide accumulation in the stomatal guard cells when compared to their wild type counterparts. They also showed significantly enhanced fresh weight, relative water content, dry weight under stress. Treatment with sodium chloride resulted in enhanced expression of genes for Delta-pyrroline-5-carboxylase synthetase in leaves, and sucrose phosphate synthase, sucrose synthase and cellulose synthase A in the leaves and fibers of transgenic plants. The transgenic plants maintained normal seed development, fiber quality and cellulose content under stress.
Plant Mol Biol 2010 Jun
PMID:Constitutive expression of mustard annexin, AnnBj1 enhances abiotic stress tolerance and fiber quality in cotton under stress. 2014 50

Transfer cells (TCs) are specialized cells exhibiting invaginated wall ingrowths (WIs), thereby amplifying their plasma membrane surface area (PMSA) and hence the capacity to transport nutrients. However, it remains unknown as to whether TCs play a role in biomass yield increase during evolution or domestication. Here, we examine this issue from a comparative evolutionary perspective. The cultivated tetraploid AD genome species of cotton and its A and D genome diploid progenitors displayed high, medium, and low seed and fiber biomass yield, respectively. In all three species, cells of the innermost layer of the seed coat juxtaposed to the filial tissues trans-differentiated to a TC morphology. Electron microscopic analyses revealed that these TCs are characterized by sequential formation of flange and reticulate WIs during the phase of rapid increase in seed biomass. Significantly, TCs from the tetraploid species developed substantially more flange and reticulate WIs and exhibited a higher degree of reticulate WI formation than their progenitors. Consequently, the estimated PMSA of TCs of the tetraploid species was about 4 and 70 times higher than that of TCs of the A and D genome progenitors, respectively, which correlates positively with seed and fiber biomass yield. Further, TCs with extensive WIs in the tetraploid species had much stronger expression of sucrose synthase, a key enzyme involved in TC WI formation and function, than those from the A and D progenitors. The analyses provide a set of novel evidence that the development of TC WIs may play an important role in the increase of seed and fiber biomass yield through polyploidization during evolution.
Mol Plant 2010 Nov
PMID:Evidence for the role of transfer cells in the evolutionary increase in seed and fiber biomass yield in cotton. 2086 53


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