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Barley stripe mosaic virus (BSMV)-based virus induced gene silencing (VIGS) is an effective strategy for rapid determination of functional genes in wheat plants. ERECTA genes are reported to regulate stomatal pattern of plants, and manipulation of TaERECTA (a homologue of ERECTA in bread wheat) is a potential route for investigating stomatal development. Here, the leucine-rich repeat domains (LRRs) and transmembrane domains of TaERECTA were selected to gain BSMV:ER-LR and BSMV:ER-TM constructs, respectively, targeting TaERECTA for silencing in wheat cultivars ‘Bobwhite’ and ‘Cadenza’, to identify the function of TaERECTA on stomatal patterns. The results showed that reduced expression of TaERECTA caused an increased stomatal and epidermal cell density by average 13.5% and 3.3%, respectively, due to the significantly reduced size of leaf epidermal and stomatal cells, and this led to an increase in stomatal conductance. These suggest that modulation of TaERECTA offers further opportunities in stomatal engineering for the adaptation of photosynthesis in wheat.
Gibberellins (GAs) are a class of plant hormones that play important roles in diverse aspects during plant growth and development. A series of GA synthesis and metabolism genes have been reported or proved to have essential functions in different plant species, while a small number of GA 2-oxidase genes have been cloned or reported in wheat. Previous studies have provided some important findings on the process of GA biosynthesis and the enzymes involved in its related pathways. These may facilitate understanding of the complicated process underlying GA synthesis and metabolism in wheat. In this study, GA 2-oxidase genes TaGA2ox1-1, TaGA2ox1-2, TaGA2ox1-3, TaGA2ox1-4, TaGA2ox1-5, and TaGA2ox1-6 were identified and further overexpressed in rice plants to investigate their functions in GA biosynthesis and signaling pathway. Results showed overexpression of GA 2-oxidase genes in rice disrupted the GA metabolic pathways and induced catalytic responses and regulated other GA biosynthesis and signaling pathway genes, which further leading to GA signaling disorders and diversity in phenotypic changes in rice plants.
Leaf senescence is a notably important trait that limits the yield and biomass accumulation of agronomic crops. Therefore, determining the chromosomal position of the expression sequence tags (ESTs) that are associated with leaf senescence is notably interesting in the manipulation of leaf senescence for crop improvement. A total of 32 ESTs that were previously identified during the delaying leaf senescence stage in the stay-green wheat cultivar CN17 were mapped to 42 chromosomes, a chloroplast, a mitochondrion, and a ribosome using in silico mapping. Then, we developed 19 pairs of primers based on these sequences and used them to determine the polymorphisms between the stay-green cultivars (CN12, CN17, and CN18) and the control cultivar MY11. Among the 19 pairs of primers, 5 pairs produced polymorphisms between the stay-green cultivar and the non-stay-green control. Further studies of Chinese Spring nullisomic-tetrasomics show that JK738991 is mapped to 3B, JK738983 is mapped to 5D, and JK738989 is mapped to 2A, 4A, and 3D. The other two ESTs, JK738994 and JK739003, were not assigned to a chromosome using the Chinese Spring nullisomic-tetrasomics, which indicates that these ESTs may be derived from rye DNA in the wide cross. In particular, the ESTs that produce polymorphisms are notably useful in identifying the stay-green cultivar using molecular marker-assisted selection. The results also suggest that the in silico mapping data, even from a comparison genomic analysis based on the homogeneous comparison, are useful at some points, but the data were not always reliable, which requires further investigation using experimental methods.
A reborn interest has occurred during the last decade toward wheat landraces for broadening genetic basis of modern wheat cultivars. The investigation of molecular traits within and between existing landraces of wheat can help scientists to develop appropriate strategies for their efficient maintenance and exploitation. The present study dealt with the gliadin characterization of forty-seven wheat landraces collected from wheat mainly planted areas of China, each of which was represented by a sample of at least 43 individuals. Twelve accessions selected on the basis of gliadin analysis were investigated further using 21 SSR markers. The results proved that landraces of wheat are a mixture of variable individuals genetically distinguishable from each other. Twelve of the analyzed 47 accessions were observed to be homogeneous, while 35 (74.5%) of them were heterogeneous in their gliadin composition. In total, 122 gliadin pattern were observed. On average, 10.1% (Gst) of the total variation arose from differentiation among regions, and 89.9% was attributed to within-region variation. Furthermore, nineteen of the 21 SSR markers were polymorphic across all the populations. The total number of the amplified DNA products was 110, with a mean of 6.11 alleles per locus. The values of genetic diversity within each landrace population varied from 0.006 to 0.351. Implications for the management of this valuable genetic resource are discussed.
Youzimai is a widespread wheat landrace and has been used extensively in breeding programs in China. In order to assess the genetic variation between and within Youzimai accessions, samples of 31 landrace accessions of wheat, all called ‘Youzimai’, were collected from 6 geographic regions in China and evaluated using morphological traits, seedling resistance to powdery mildew, gliadin and microsatellite markers. Typical differences among accessions were observed in morphological characteristics. Forty-five (58.4%) of 77 assayed SSR markers showed polymorphism over the entire collection and total 226 alleles were identified with an average of 5.02 alleles per locus. SSR data indicated that the accessions from Hebei province were the most diverse, as evidenced by greatest number of region-specific alleles and highest diversity index. These accessions, therefore, probably experienced the most substantial morphological and molecular evolution as a result of various natural and anthropomorphic influences. On the other hand, differentiation in gliadin phenotypes was found among seeds within 80.6% of total accessions and average 61.5% of entire collections showed heterogeneous and comprised resistant plants in reaction to powdery mildew, suggesting the presence of a wide diversity within the wheat landrace. By developing an intimate knowledge of the available wheat genotypes, appropriate selections can be made for commercial application in order to conserve and exploit the diversity of the wheat landraces.
As one of the world’s earliest domesticated crops, barley is a model species for the study of evolution and domestication. Domestication is an evolutionary process whereby a population adapts, through selection; to new environments created by human cultivation. We describe the genome-scanning of molecular diversity to assess the evolution of barley in the Tibetan Plateau. We used 667 Diversity Arrays Technology (DArT) markers to genotype 185 barley landraces and wild barley accessions from the Tibetan Plateau. Genetic diversity in wild barley was greater than in landraces at both genome and chromosome levels, except for chromosome 3H. Landraces and wild barley accessions were clearly differentiated genetically, but a limited degree of introgression was still evident. Significant differences in diversity between barley subspecies at the chromosome level were observed for genes known to be related to physiological and phenotypical traits, disease resistance, abiotic stress tolerance, malting quality and agronomic traits. Selection on the genome of six-rowed naked barley has shown clear multiple targets related to both its specific end-use and the extreme environment in Tibet. Our data provide a platform to identify the genes and genetic mechanisms that underlie phenotypic changes, and provide lists of candidate domestication genes for modified breeding strategies.
Seven Glu-A1 m allelic variants of the Glu-A1 m x genes in Triticum monococcum ssp. monococcum, designated as 1Ax2.1 a , 1Ax2.1 b , 1Ax2.1 c , 1Ax2.1 d , 1Ax2.1 e , 1Ax2.1 f , and 1Ax2.1 g were characterized. Their authenticity was confirmed by successful expression of the coding regions in E. coli, and except for the 1Ax2.1 a with the presence of internal stop codons at position of 313 aa, all correspond to the subunit in seeds. However, all the active six genes had a same DNA size although their encoding subunits showed different molecular weight. Our study indicated that amino acid residue substitutions rather than previously frequently reported insertions/deletions played an important role on the subunit evolution of these Glu-A1 m x alleles. Since variation in the Glu-A1x locus in common wheat is rare, these novel genes at the Glu-A1 m x can be used as candidate genes for further wheat quality improvement.
Nitrogen (N) is an important nutrient for plant growth and yield production, and rice grown in paddy soil mainly uses ammonium (NH4 +) as its N source. Previous studies have shown that N status is tightly connected to plant defense; however, the roles of NH4 + uptake and assimilation in rice sheath blight disease response have not been studied previously. Here, we analyzed the effects of different N sources on plant defense against Rhizoctonia solani. The results indicated that rice plants grown in N-free conditions had higher resistance to sheath blight than those grown under N conditions. In greater detail, rice plants cultured with glutamine as the sole N source were more susceptible to sheath blight disease compared to the groups using NH4 + and nitrate (NO3 –) as sole N sources. N deficiency severely inhibited plant growth; therefore, ammonium transporter 1;2 overexpressors (AMT1;2 OXs) were generated to test their growth and defense ability under low N conditions. AMT1;2 OXs increased N use efficiency and exhibited less susceptible symptoms to R. solani and highly induced the expression of PBZ1 compared to the wild-type controls upon infection of R. solani. Furthermore, the glutamine synthetase 1;1 (GS1;1) mutant (gs1;1) was more susceptible to R. solani infection than the wild-type control, and the genetic combination of AMT1;2 OX and gs1;1 revealed that AMT1;2 OX was less susceptible to R. solani and required GS1;1 activity. In addition, cellular NH4 + content was higher in AMT1;2 OX and gs1;1 plants, indicating that NH4 + was not directly controlling plant defense. In conclusion, the present study showed that the activation of NH4 + uptake and assimilation were required for rice resistance against sheath blight disease.
Higher plant population and nitrogen management is an adopted approach for improving crop productivity from limited land resources. Moreover, higher plant density and nitrogen regimes may increase the risk of stalk lodging, which is a consequence of complex interplant competition of individual organs. Here, we aimed to investigate the dynamic change in morphology, chemical compositions and lignin promoting enzymes of the second basal inter-nodes altering lodging risk controlled by planting density and nitrogen levels. A field trial was conducted at the Mengcheng research station (33°9′44″N, 116°32′56″E), Huaibei plain, Anhui province, China. A randomized complete block design was adopted, in which four plant densities, i.e., 180, 240, 300, and 360 × 104 ha−1 and four N levels, i.e., 0, 180, 240, and 300 kg ha−1 were studied. The two popular wheat varieties AnNong0711 and YanNong19 were cultivated. Results revealed that the culm lodging resistance (CLRI) index of the second basal internodes was positively and significantly correlated with light interception, lignin and cellulose content. The lignin and cellulose contents were significantly and positive correlated to light interception. The increased planting density and nitrogen levels declined the lignin and its related enzymes activities. The variety AnNong0711 showed more resistive response to lodging compared to YanNong19. Overall our study found that increased planting densities and nitrogen regimes resulted in poor physical strength and enzymatic activity which enhanced lodging risk in wheat varieties. The current study demonstrated that stem bending strength of the basal internode was significantly positive correlated to grains per spike. The thousand grain weight and grain yield had a positive and significant relationship with stem bending strength of the basal internode. The results suggested that the variety YanNong19 produces higher grain yield (9298 kg ha−1) at density 240 × 104 plants ha−1, and 180 kg ha−1 nitrogen, while AnNong0711 produced higher grain yield (10178.86 kg ha−1) at density 240 × 104 plants ha−1 and with 240 kg ha−1 nitrogen. Moreover, this combination of nitrogen and planting density enhanced the grain yield with better lodging resistance.
The forward and reverse cDNA subtractive libraries before and after the toxic effect of α-amanitin were constructed by suppression subtractive hybridization and randomly selected clones from each subtractive library were screened by PCR and dot blot hybridization. A total of 85 genes with altered expression were finally identified, with 41 genes from the forward library and 44 from the reverse library. Subsequently, the antagonistic effects of candidate traditional Chinese medicines were evaluated based on the genetic transcription levels of the genes with significant altered expression, including Catnβ, Flt3-L, IL-7r and Rpo2-4. The results indicated that Silybum marianum (L.) Gaert and Ganoderma lucidum had significant down-regulated effects on the transcription level of Catnβ that was up-regulated by α-amanitin, and the two herbs also up-regulated the transcription levels of Flt3-L and Rpo2-4. Silybum marianum (L.) had significant up-regulated effects on the IL-7r that was down-regulated by α-amanitin. These preliminary studies suggested that Silybum marianum (L.) and Ganoderma lucidum were effective antagonists against the toxicity of α-amanitin.
