Increased gibberellin contents contribute to accelerated growth and development of transgenic tobacco overexpressing a wheat ubiquitin gene

Key message

Overexpressing TaUb2 promoted stem growth and resulted in early flowering in transgenic tobacco plants. Ubiquitin are involved in the production, metabolism and proper function of gibberellin.

Abstract

The ubiquitin–26S proteasome system (UPS), in which ubiquitin (Ub) functions as a marker, is a post-translational regulatory system that plays a prominent role in various biological processes. To investigate the impact of different Ub levels on plant growth and development, transgenic tobacco (Nicotiana tabacum L.) plants were engineered to express an Ub gene (TaUb2) from wheat (Triticum aestivum L.) under the control of cauliflower mosaic virus 35S promoter. Transgenic tobacco plants overexpressing TaUb2 demonstrated an accelerated growth rate at early stage and an early flowering phenotype in development. The preceding expression of MADS-box genes also corresponded to the accelerated developmental phenotypes of the transgenic tobacco plants compared to that of wild-type (WT). Total gibberellin (GA) and active GA contents in transgenic tobacco plants were higher than those in WT at the corresponding developmental stages, and some GA metabolism genes were upregulated. Treatment with GA₃ conferred a similarly accelerated grown rate in WT plants to that of transgenic tobacco plants, while growth was inhibited when transgenic tobacco plants were treated with a GA biosynthesis inhibitor. Thus, the results suggest that Ub are involved in the production, metabolism and proper function of GA, which is important in the regulation of plant growth and development.     

Nutrient partitioning and grain yield of TaNAM-RNAi wheat under abiotic stress

Aims

Decreased expression of TaNAM genes by RNAi results in delayed senescence and decreased grain protein, iron, and zinc concentrations. Here, we determined whether NAM expression level alters onset of senescence under stress conditions, whether delayed senescence in the TaNAM-RNAi line resulted in improved tolerance to post-anthesis abiotic stress, and determined the effects of post-anthesis abiotic stress on N and mineral remobilization and partitioning to grain.

Methods

Greenhouse-grown WT and TaNAM-RNAi wheat were characterized in two studies:three levels of N fertility or water limitation during grain fill. Studies were conducted under both optimal and heat stress temperatures. Senescence onset was determined by monitoring flag leaf chlorophyll.

Results

Under optimal tempertures, TaNAM-RNAi plants had a yield advantage at lower N. TaNAM-RNAi plants had delayed senescence relative to the WT and lower grain protein and mineral concentrations, N remobilization efficiency, and partitioning of N and most minerals to grain.

Conclusions

Nutritional quality of TaNAM-RNAi grain was consistently lower than WT. Delayed senescence of TaNAM-RNAi plants provided a yield advantage under optimal temperatures but not under water or heat stress. Discovery of specific NAM protein targets may allow separation of the delayed senescence and nutrient partitioning traits, which could be used for improvement of wheat.     

Genome-wide patterns of genetic variation in sweet and grain sorghum (Sorghum bicolor)

Background

Sorghum (Sorghum bicolor) is globally produced as a source of food, feed, fiber and fuel. Grain and sweet sorghums differ in a number of important traits, including stem sugar and juice accumulation, plant height as well as grain and biomass production. The first whole genome sequence of a grain sorghum is available, but additional genome sequences are required to study genome-wide and intraspecific variation for dissecting the genetic basis of these important traits and for tailor-designed breeding of this important C₄ crop.

Results

We resequenced two sweet and one grain sorghum inbred lines, and identified a set of nearly 1,500 genes differentiating sweet and grain sorghum. These genes fall into ten major metabolic pathways involved in sugar and starch metabolisms, lignin and coumarin biosynthesis, nucleic acid metabolism, stress responses and DNA damage repair. In addition, we uncovered 1,057,018 SNPs, 99,948 indels of 1 to 10 bp in length and 16,487 presence/absence variations as well as 17,111 copy number variations. The majority of the large-effect SNPs, indels and presence/absence variations resided in the genes containing leucine rich repeats, PPR repeats and disease resistance R genes possessing diverse biological functions or under diversifying selection, but were absent in genes that are essential for life.

Conclusions

This is a first report of the identification of genome-wide patterns of genetic variation in sorghum. High-density SNP and indel markers reported here will be a valuable resource for future gene-phenotype studies and the molecular breeding of this important crop and related species.     

Expression of sorghum gene <Emphasis Type="Italic">SbSGL</Emphasis> enhances grain length and weight in rice

Grain weight is a major determining factor of rice (Oryza sativa L.) yield and the comprehensive embodiment of grain length, width, and thickness. Here, we describe the molecular and functional characterization of SbSGL (Sorghum bicolor L. stress tolerance and grain length), a sorghum gene that encodes a putative member of the DUF1645 protein family of unknown function. Expression of SbSGL in rice promoted cell division and grain filling, which affected an array of traits of rice, including grain length, grain weight, and seed setting rate. Expression of SbSGL also affected the expression of genes related to the plant cell cycle and grain size.     

Impact of the 7-bp deletion in <Emphasis Type="Italic">HvGA20ox2</Emphasis> gene on agronomic important traits in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Background

Alike to Reduced height-1 (Rht-1) genes in wheat and the semi dwarfing (sd-1) gene in rice, the sdw1/denso locus involved in the metabolism of the GA, was designated as the ‘Green Revolution’ gene in barley. The recent molecular characterization of the candidate gene HvGA20ox2 for sdw1/denso locus allows to estimate the impact of the functional polymorphism of this gene on the variation of agronomically important traits in barley.

Results

We investigated the effect of the 7-bp deletion in exon 1 of HvGA20ox2 gene (sdw1.d mutation) on the variation of yield-related and malting quality traits in the population of DHLs derived from cross of medium tall barley Morex and semi-dwarf barley Barke. Segregation of plant height, flowering time, thousand grain weight, grain protein content and grain starch was evaluated in two diverse environments separated from one another by 15° of latitude. The 7-bp deletion in HvGA20ox2 gene reduced plant height by approximately 13 cm and delayed flowering time by 3–5 days in the barley segregating DHLs population independently on environmental cue. On other hand, the sdw1.d mutation did not affect significantly either grain quality traits (protein and starch content) or thousand grain weight.

Conclusions

The beneficial effect of the sdw1.d allele could be associated in barley with lodging resistance and extended period of vegetative growth allowing to accumulate additional biomass that supports higher yield in certain environments. However, no direct effect of the sdw1.d mutation on thousand grain weight or grain quality traits in barley was detected.

     

Altered gibberellin content affects growth and development in transgenic tobacco lines overexpressing a wheat gene encoding F-box protein

In a previous study, we have identified and characterized gene from wheat (Triticum aestivum L.) encoding F-box protein and named it TaFBA. In this paper, transgenic tobacco (Nicotiana tabacum L.) plants overexpressing TaFBA1 displayed accelerated growth early, but the rate slowed gradually at later stages of growth, and the mature transgenic plants were even shorter in stature and flowered later than did the wild type (WT). Treatment with gibberellin (GA) conferred an accelerated growth rate to the transgenic tobacco plants at later stages, similar to that of WT, whereas growth was inhibited more seriously in WT than in transgenic tobacco when plants were treated with a GA biosynthesis inhibitor. The content of GA in transgenic tobacco plants was higher at early developmental stages, but it was lower at later growth stages than in WT. Some GA biosynthesis genes were down regulated, which was accompanied with elevated expression of a GA catabolism gene. Thus, our results suggest that TaFBA1 is possibly involved in the regulation of plant growth and development, and that it may be related to the production, metabolism, and proper function of GA.     

Fine mapping and identification of a novel locus <Emphasis Type="Italic">qGL12.2</Emphasis> control grain length in wild rice (<Emphasis Type="Italic">Oryza rufipogon</Emphasis> Griff.)

Key message

A wild rice QTL qGL12.2 for grain length was fine mapped to an 82-kb interval in chromosome 12 containing six candidate genes and none was reported previously.

Abstract

Grain length is an important trait for yield and commercial value in rice. Wild rice seeds have a very slender shape and have many desirable genes that have been lost in cultivated rice during domestication. In this study, we identified a quantitative trait locus, qGL12.2, which controls grain length in wild rice. First, a wild rice chromosome segment substitution line, CSSL41, was selected that has longer glume and grains than does the Oryza sativa indica cultivar, 9311. Next, an F₂ population was constructed from a cross between CSSL41 and 9311. Using the next-generation sequencing combined with bulked-segregant analysis and F₃ recombinants analysis, qGL12.2 was finally fine mapped to an 82-kb interval in chromosome 12. Six candidate genes were found, and no reported grain length genes were found in this interval. Using scanning electron microscopy, we found that CSSL41 cells are significantly longer than those of 9311, but there is no difference in cell widths. These data suggest that qGL12.2 is a novel gene that controls grain cell length in wild rice. Our study provides a new genetic resource for rice breeding and a starting point for functional characterization of the wild rice GL gene.

     

Rice inositol polyphosphate kinase gene (<Emphasis Type="Italic">OsIPK2</Emphasis>), a putative new player of gibberellic acid signaling,involves in modulation of shoot elongation and fertility

Gibberellic acid (GA) is an important plant hormone mediating plant growth and development throughout the life span. Although many GA biosynthesis genes and signaling components have been revealed, the signal transduction mechanisms from GA perception to physiological actions are still largely unclear. In this study, we investigated the functions of a rice (Oryza sativa) inositol polyphosphate kinase gene (OsIPK2) in rice growth and development, showing that OsIPK2 is a putative new player in GA signaling. OsIPK2 is widely expressed in rice with high accumulation in tender and rapidly dividing tissues. The OsIPK2 protein is mainly localized in the nucleus and plasma membrane. To study the biological roles of OsIPK2 in rice, RNA interference and overexpression transgenic plants were generated. OsIPK2 antisense plants exhibited taller seedling height and lower fertility rate than the wild type, while overexpression lines showed reduced plant height. Microarray and qRT-PCR assays showed that expression levels of several GA-related genes were altered in transgenic plants. Besides, down-regulation of OsIPK2 resulted in hypersensitivity to paclobutrazol (PAC), a GA biosynthesis inhibitor. We also described that the expression of OsIPK2 could be either induced by GA or repressed by PAC. Taken together, these findings suggested that OsIPK2 is likely a negative regulator of GA signaling and involves in modulating shoot elongation and fertility.     

Effects of nitrogen fertilization and root interaction on the agronomic traits of intercropped maize, and the quantity of microorganisms and activity of enzymes in the rhizosphere

Aims

To elucidate the mechanisms of the beneficial effects of below-ground root interactions in maize plus legume intercropping system,

Methods

A pot experiment was conducted using root separation techniques.

Results

It is shown that root interaction and nitrogen fertilization increased chlorophyll content and improved plant characteristics of maize, and the effect of root interaction was significant (p<0.05). Compared to a full root separation treatment, no root separation increased the leaf and grain nitrogen contents, and economic and biological yields per maize plant by 9.3? %, 6.0? %, 14.0? %, and 6.5? %, respectively. Root interaction and nitrogen fertilization enhanced the numbers of bacteria, fungi, actinomycetes and Azotobacteria and the activities of urease, invertase, acid-phosphatase and protease in soil. Correlation analyses revealed that the quantity of microorganisms and the activity of the aforementioned enzymes were all positively or significantly (p<0.05) positively correlated with chlorophyll content, plant height and economic and biological yields per maize plant.

Conclusions

The findings demonstrate that root interactions are important in improving the soil micro-ecological environment, increasing microbial quantity and enzyme activity in soil, and enhancing crop yield.     

Phosphorus accumulation in grains of japonica rice as affected by nitrogen fertilizer

Background and aims

This study aims to investigate the effect of nitrogen (N) on grain phosphorus (P) accumulation in japonica rice.

Methods

Six cultivars with contrasting agronomic traits were grown for 3 years (from 2008 to 2010) of field experiments under seven N treatments and 1 year (in 2010) of pot experiments with five N treatments to study the effect of N on grain phosphorus accumulation and to explore its physiological foundation.

Results

Grain total P and phytic acid concentration showed a clearly decreasing trend as N rate increased for both field and pot experiments. Pot experiment revealed that application of N increase plant biomass, but tended to lower plant P uptake, especially for the split topdressing treatments. Both harvest index (HI) and P harvest index (PHI) increased with N rate, but PHI was consistently higher than HI, indicating the larger proportion of P translocation to grain than that of dry matter by N. Further, ratio of PHI/HI differed significantly among genotypes, but was stable across contrasting N treatments.

Conclusions

The combination of decreased plant P uptake and dilution effect of increased grain yield by N is proposed as underlying mechanism of the decreased grain P concentration by high N.     

GmPAP4, a novel purple acid phosphatase gene isolated from soybean (Glycine max), enhanced extracellular phytate utilization in Arabidopsis thaliana

Key message

GmPAP4 , a novel plant PAP gene in soybean, has phytase activity. Over-expressing GmPAP4 can enhance Arabidopsis growth when phytate is the sole P source in culture.

Abstract

Phosphorus (P) is an important macronutrient for plant growth and development. However, most of the total P in soils is fixed into organic phosphate (Po). Purple acid phosphatase (PAP) can hydrolyze Po in the soil to liberate inorganic phosphate and enhance plant P utilization. We isolated a novel PAP gene, GmPAP4, from soybean (Glycine max). It had an open reading frame of 1,329 bp, encoding 442 amino acid residues. Sequence alignment and phylogenetics analysis indicated that GmPAP4 was similar to other plant PAPs with large molecular masses. Quantitative real-time PCR analysis showed that the induced expression of GmPAP4 was greater in P-efficient genotype Zhonghuang15 (ZH15) than in P-inefficient genotype Niumaohuang (NMH) during the periods of flowering (28–35 days post phytate stress; DPP) and pod formation (49–63 DPP). Moreover, peak expression, at 63 DPP, was about 3-fold higher in ‘ZH15’ than in ‘NMH’. Sub-cellular localization showed that GmPAP4 might be on plasma membrane or in cytoplasm. Over-expressing GmPAP4 in Arabidopsis resulted in significant rises in P acquisition and utilization compared with the wild-type (WT). Under phytate condition, transgenic Arabidopsis plants showed increases of approximately 132.7 % in dry weight and 162.6 % in shoot P content compared with the WT. Furthermore, when phytate was added as the sole P source in cultures, the activity of acid phosphatase was significantly higher in transgenic plants. Therefore, GmPAP4 is a novel PAP gene that functions in plant’s utilization of organic phosphate especially under phytate condition.     

Enhanced tolerance to drought and salt stresses in transgenic faba bean (Vicia faba L.) plants by heterologous expression of the PR10a gene from potato

Key message

We report for the first time that expression of potato PR10a gene in faba bean causes enhanced tolerance to drought and salinity.

Abstract

Grain legumes such as soybean (Glycine max L. Merrill), pea (Pisum sativum L.) and faba bean (Vicia faba L.) are staple sources of protein for human and animal nutrition. Among grain legumes, faba bean is particularly sensitive to abiotic stress (in particular osmotic stress due to lack of water or enhanced soil salinity) and often suffers from severe yield losses. Many stress responsive genes have been reported with an effect on improving stress tolerance in model plants. Pathogenesis-related proteins are expressed by all plants in response to pathogen infection and, in many cases, in response to abiotic stresses as well. The PR10a gene isolated from the potato cultivar Desiree was selected for this study due to its role in enhancing salt and/or drought tolerance in potato, and transferred into faba bean cultivar Tattoo by Agrobacterium tumefaciens-mediated transformation system based upon direct shoot regeneration after transformation of meristematic cells derived from embryo axes. The transgene was under the control of the constitutive mannopine synthase promoter (p-MAS) in a dicistronic binary vector, which also contained luciferase (Luc) gene as scorable marker linked by internal ribosome entry site elements. Fertile transgenic faba bean plants were recovered. Inheritance and expression of the foreign genes were demonstrated by PCR, RT-PCR, Southern blot and monitoring of Luciferase activity. Under drought condition, after withholding water for 3 weeks, the leaves of transgenic plants were still green, while non-transgenic plants (WT) wilted and turned brown. Twenty-four hours after re-watering, the leaves of transgenic plants remained green, while WT plants did not recover. Moreover, the transgenic lines displayed higher tolerance to NaCl stress. Our results suggested that introducing a novel PR10a gene into faba bean could be a promising approach to improve its drought and salt tolerance ability, and that MAS promoter is not only constitutive, but also wound-, auxin/cytokinin- as well as stress-inducible.     

Contrasting effects of dwarfing alleles and nitrogen availability on mineral concentrations in wheat grain

Background and aim

Concentrations of essential minerals in plant foods may have declined in modern high-yielding cultivars grown with large applications of nitrogen fertilizer (N). We investigated the effect of dwarfing alleles and N rate on mineral concentrations in wheat.

Methods

Gibberellin (GA)-insensitive reduced height (Rht) alleles were compared in near isogenic wheat lines. Two field experiments comprised factorial combinations of wheat variety backgrounds, alleles at the Rht-B1 locus (rht-B1a, Rht-B1b, Rht-B1c), and different N rates. A glasshouse experiment also included Rht-D1b and Rht-B1b+D1b in one background.

Results

In the field, depending on season, Rht-B1b increased crop biomass, dry matter (DM) harvest index, grain yield, and the economically-optimal N rate (N _opt ). Rht-B1b did not increase uptake of Cu, Fe, Mg or Zn so these minerals were diluted in grain. Nitrogen increased DM yield and mineral uptake so grain concentrations were increased (Fe in both seasons; Cu, Mg and Zn in one season). Rht-B1b reduced mineral concentrations at N _opt in the most N responsive season. In the glasshouse experiment, grain yield was reduced, and mineral concentrations increased, with Rht allele addition.

Conclusion

Effects of Rht alleles on Fe, Zn, Cu and Mg concentrations in wheat grain are mostly due to their effects on DM, rather than of GA-insensitivity on N _opt or mineral uptake. Increased N requirement in semi-dwarf varieties partly offsets this dilution effect.