Identification of Reference Genes for RT-qPCR Data Normalization in Cannabis sativa Stem Tissues

Gene expression profiling via quantitative real-time PCR is a robust technique widely used in the life sciences to compare gene expression patterns in, e.g., different tissues, growth conditions, or after specific treatments. In the field of plant science, real-time PCR is the gold standard to study the dynamics of gene expression and is used to validate the results generated with high throughput techniques, e.g., RNA-Seq. An accurate relative quantification of gene expression relies on the identification of appropriate reference genes, that need to be determined for each experimental set-up used and plant tissue studied. Here, we identify suitable reference genes for expression profiling in stems of textile hemp (Cannabis sativa L.), whose tissues (isolated bast fibres and core) are characterized by remarkable differences in cell wall composition. We additionally validate the reference genes by analysing the expression of putative candidates involved in the non-oxidative phase of the pentose phosphate pathway and in the first step of the shikimate pathway. The goal is to describe the possible regulation pattern of some genes involved in the provision of the precursors needed for lignin biosynthesis in the different hemp stem tissues. The results here shown are useful to design future studies focused on gene expression analyses in hemp.     

Metabolomics Mechanism and Lignin Response to Laxogenin C,a Natural Regulator of Plants Growth

Laxogenin C (LGC) is a natural spirostanol deriving from plant hormone which has shown growing regulation similar to those of brassinosteroids. In the present study, LGC showed a promoting effect on tomato seed germination and seedling growth in a dose-dependent manner. We applied LC-MS/MS to investigate metabolome variations in the tomato treated with LGC, which revealed 10 differential metabolites (DMs) related to KEGG metabolites, associated with low and high doses of LGC. Enrichment and pathway mapping based on the KEGG database indicated that LGC regulated expressions of 2-hydroxycinnamic acid and l-phenylalanine to interfere with phenylalanine metabolism and phenylpropanoids biosynthesis. The two pathways are closely related to plant growth and lignin formation. In our further phenotypic verification, LGC was confirmed to affect seedling lignification and related phenylpropanoids, trans-ferulic acid and l-phenylalanine levels. These findings provided a metabolomic aspect on the plant hormone derivates and revealed the affected metabolites. Elucidating their regulation mechanisms can contribute to the development of sustainable agriculture. Further studies on agrichemical development would provide eco-friendly and efficient regulators for plant growth control and quality improvement.     

壳聚糖-乳清分离蛋白复合膜对鲜切雷竹笋木质化和品质的影响

采用壳聚糖-乳清分离蛋白复合膜（chitosan-whey protein isolate composite film,CWF）涂膜1、2、3 次及去离子水、乙酸处理鲜切雷竹笋,于4 ℃冷藏15 d,以贮藏过程雷竹笋的苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）活力、丙二醛（MDA）、多酚、木质素含量、菌落总数等为指标,研究CWF对鲜切雷竹笋木质化和品质劣变以及涂膜次数对保鲜效果的影响。结果显示,乙酸对鲜切雷竹笋的木质化和微生物有一定的抑制作用,但影响不显著;CWF涂膜可以显著降低鲜切雷竹笋表面的微生物数量,抑制PAL和POD活力,降低木质素、MDA含量,延长鲜切雷竹笋的货架期。随着CWF涂膜次数的增加,保鲜效果更好,考虑到生产成本选择涂膜2 次。     

不同贮藏条件对山竹果皮木质化效应及抑菌活性影响

以海南山竹为材料,研究了不同贮藏条件对山竹果皮木质化及抑菌活性影响。结果表明:随着贮藏时间的延长,山竹果壳木质素沉积指标木质素含量和粗纤维含量均呈上升趋势。贮藏末期,常温处理的木质素含量比同期壳聚糖、海藻酸钠和低温处理的木质素含量分别增加6.0%、2.8%和14.3%。木质化底物总酚含量呈先升后降趋势,在第6天达到峰值,为33.033 mgGAE/g(鲜重),抑菌活性表明,在贮藏期间,山竹果皮抑菌活性因受木质化产物和多酚转化产物的影响有不同的变化趋势,低温和涂膜有助于抑菌活性稳态化。     

Genome-Wide Identification of Switchgrass Laccases Involved in Lignin Biosynthesis and Heavy-Metal Responses

Plant laccase genes belong to a multigene family, play key roles in lignin polymerization, and participate in the resistance of plants to biotic and abiotic stresses. Switchgrass is an important resource for forage and bioenergy production, yet information about the switchgrass laccase gene family is scarce. Using bioinformatic approaches, a genome-wide analysis of the laccase multigene family in switchgrass was carried out in this study. In total, 49 laccase genes (PvLac1 to PvLac49) were identified; these can be divided into five subclades, and 20 of them were identified as targets of miR397. The tandem and segmental duplication of laccase genes on Chr05 and Chr08 contributed to the expansion of the laccase family. The laccase proteins shared conserved signature sequences but displayed relatively low sequence similarity, indicating the potential functional diversity of switchgrass laccases. Switchgrass laccases exhibited distinct tissue/organ expression patterns, revealing that some laccases might be involved in the lignification process during stem development. All five of the laccase isoforms selected from different subclades responded to heavy metal. The immediate response of lignin-related laccases, as well as the delayed response of low-abundance laccases, to heavy-metal treatment shed light on the multiple roles of laccase isoforms in response to heavy-metal stress.     

Kinetic characterization of a basic peroxidase from garlic (Allium sativum L.) cloves

Peroxidases catalyze the reduction of H(2)O(2) by taking electrons from a variety of compounds from the secondary metabolism including flavonoids and lignin precursors. This work describes the purification and kinetic characterization of a basic peroxidase from garlic cloves using quercetin and p-coumaric acid, flavonoid and phenolic compounds found in garlic cloves. The high catalytic efficiency shown by this basic peroxidase in the oxidation of quercetin at acidic pH suggests good adaptation of this enzyme, involved in quercetin catabolism in the acidic physiological pH conditions of the vacuoles, where it is presumably located. Likewise, garlic peroxidase showed similar oxidation rates for hydroxycinnamyl (p-coumaric) and sinapyl-type structures, which suggests its involvement in the cross-coupling reactions that occur in the cell wall during lignification. On the other hand, the high affinity of this enzyme for H(2)O(2) would be in accordance with the oxidation of both flavonoid and phenolic compounds to regulate H(2)O(2) levels in tissues/organelles, where this peroxidase is expressed.     

Effect of Storage on the Biochemical Structure and Processing Quality of Adzuki Bean (Vigna angularis)

Storage of adzuki beans and other pulse grains causes biochemical and physical changes that affect the hydration properties of the beans. This affects the quality of products made from the beans such as the Japanese bean paste “ann.” Storage, particularly under unfavourable conditions, leads to the “hard shell” phenomenon, where beans fail to imbibe water when soaked and remain hard, and the “hard-to-cook” phenomenon where the seeds hydrate normally, but the cotyledon fails to hydrate and soften during cooking. The hard shell phenomenon is attributable to impermeability of the seed coat to water, which is due to biochemical changes in the seed coat, such as the formation of protein-tannin complexes, and biophysical changes such as reduction in size or closure of the straphiole aperture in the hilum area—the main area for water entry into the adzuki bean. The hard-to-cook phenomenon is due to changes in the cotyledon tissue, which include formation of insoluble pectinates, lignification of the cell wall and middle lamella, interaction of condensed tannins with proteins and starch, and changes to the structure and functionality of the cellular proteins and starch.