Astragalus membranaceus, also known as Huangqi in China, is one of the most widely used medicinal herbs in Traditional Chinese Medicine. Traditional Chinese Medicine formulations from Astragalus membranaceus have been used to treat a wide range of illnesses, such as cardiovascular disease, type 2 diabetes, nephritis and cancers. Pharmacological studies have shown that immunomodulating, anti-hyperglycemic, anti-inflammatory, antioxidant and antiviral activities exist in the extract of Astragalus membranaceus. Therefore, characterising the biosynthesis of bioactive compounds in Astragalus membranaceus, such as Astragalosides, Calycosin and Calycosin-7-O-β-d-glucoside, is of particular importance for further genetic studies of Astragalus membranaceus. In this study, we reconstructed the Astragalus membranaceus full-length transcriptomes from leaf and root tissues using PacBio Iso-Seq long reads. We identified 27 975 and 22 343 full-length unique transcript models in each tissue respectively. Compared with previous studies that used short read sequencing, our reconstructed transcripts are longer, and are more likely to be full-length and include numerous transcript variants. Moreover, we also re-characterised and identified potential transcript variants of genes involved in Astragalosides, Calycosin and Calycosin-7-O-β-d-glucoside biosynthesis. In conclusion, our study provides a practical pipeline to characterise the full-length transcriptome for species without a reference genome and a useful genomic resource for exploring the biosynthesis of active compounds in Astragalus membranaceus.
The movement of water into harvest-ripe grains of dormant and non-dormant genotypes of wheat (Triticum aestivum L.) was investigated using Magnetic Resonance Micro-Imaging (MRMI). Images of virtual sections, both longitudinal and transverse, throughout the grain were collected at intervals after the start of imbibition and used to reconstruct a picture of water location within the different grain tissues and changes over time. The observations were supplemented by the weighing measurements of water content and imbibition of grains in water containing I2/KI which stains starch and lipid, thereby acting as a marker for water. In closely related genotypes, with either a dormant or a non-dormant phenotype, neither the rate of increase in water content nor the pattern of water distribution within the grain was significantly different until 18 h, when germination became apparent in the non-dormant genotype. Water entered the embryo and scutellum during the very early stages of imbibition through the micropyle and by 2 h water was clearly evident in the micropyle channel. After 12 h of imbibition, embryo structures such as the coleoptile and radicle were clearly distinguished. Although water accumulated between the inner (seed coat) and outer (pericarp) layers of the coat surrounding the grain, there was no evidence for movement of water directly across the coat and into the underlying starchy endosperm.
Grain dormancy in wheat is an important component of resistance to preharvest sprouting and hence an important trait for wheat breeders. The significant influence of environment on the dormancy phenotype makes this trait an obvious target for marker-assisted-selection. Closely related breeding lines, SUN325B and QT7475, containing a major dormancy QTL derived from AUS1408 located on chromosome 4A, but substantially different in dormancy phenotype, were compared with a nondormant cultivar, Hartog, in a range of controlled environments. As temperature increased, dormancy at harvest-ripeness decreased particularly for QT7475. The dormancy phenotypes of reciprocal F 1 grains involving all possible combinations of Hartog, QT7475 and SUN325B were also compared in two environments with different temperatures. The results were consistent with the presence of QTL in addition to 4A in SUN325B, compared with QT7475, at least one of which was associated with the seed coat. Genetic analysis of a doubled haploid population derived from SUN325B 9 QT7475 identified a highly significant QTL located on chromosome 3BL, close to the expected position of the mutant allele of the red seed coat colour gene in whitegrained wheat, R-B1a. When the lines in the population were grouped according to the parental alleles at marker loci flanking the 3B QTL, the dormancy phenotype frequency distribution for the SUN325B group was shifted towards greater dormancy compared with the QT7475 group. However, significant variation for dormancy phenotype remained within each group. Lines representing the extremes of the range of phenotypes within each group maintained their relative ranking across seven environments consistent with the presence of another unidentified QTL contributing to dormancy in SUN325B.
A total of 120 Mesorhizobium strains collected from the central dry zone of Myanmar were analysed in a pot experiment to evaluate nodulation and symbiotic effectiveness (SE%) in chickpea plants. Phylogenetic analyses revealed all strains belonged to the genus Mesorhizobium according to 16-23S rDNA IGS and the majority of chickpea nodulating rhizobia in Myanmar soils were most closely related to M. gobiense, M. muleiense, M. silamurunense, M. tamadayense and M. temperatum. Around two thirds of the Myanmar strains (68%) were most closely related to Indian strain IC-2058 (CA-181), which is also most closely related to M. gobiense. There were no strains that were closely related to the cognate rhizobial species to nodulate chickpea: M. ciceri and M. mediterraneum. Strains with diverse 16S-23S rDNA IGS shared similar nodC and nifH gene sequences with chickpea symbionts. Detailed sequence analysis of the nodC and nifH found that the strains in Myanmar were somewhat divergent from the group including M. ciceri and were more closely related to M. muleiense and IC-2058. A cross continent analysis between strains isolated in Australia compared with Myanmar found that there was little overlap in species, where Australian soils were dominated with M. ciceri, M. temperatum and M. huakuii. The only co-occurring species found in both Myanmar and Australia were M. tamadayense and M. silumurunense. Continued inoculation with CC1192 may have reduced diversity of chickpea strains in Australian soils. Isolated strains in Australian and Myanmar had similar adaptive traits, which in some cases were also phylogenetically related. The genetic discrepancy between chickpea nodulating strains in Australia and Myanmar is not only due to inoculation history but to adaptation to soil conditions and crop management over a long period, and there has been virtually no loss of symbiotic efficiency over this time in strains isolated from soils in Myanmar.
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