Molecular responses reveal that two glutathione S-transferase CsGSTU8s contribute to detoxification of glyphosate in tea plants (Camellia sinensis).

Tea plant (Camellia sinensis) is an important economical crop that frequently suffers from various herbicides, especially glyphosate. However, the molecular responses and regulatory mechanisms of glyphosate stress in tea plants remain poorly understood. Here, we reported a transcriptome dataset and identified large number of differentially expressed genes (DEGs) under glyphosate exposure. Next, two glutathione S-transferase genes (CsGSTU8-1 and CsGSTU8-2) that upregulated significantly were screened as candidate genes. Tissue-specific expression patterns showed that both CsGSTU8-1 and CsGSTU8-2 had extremely high expression levels in the roots and were predominantly localized in the nucleus and plasma membrane based on subcellular localization. Both were significantly upregulated at different time points under various stressors, including drought, cold, salt, pathogen infections, and SA treatments. An enzymatic activity assay showed that CsGSTU8-1 catalyzes the conjugation of glutathione with 2,4-dinitrochlorobenzene (CDNB). Functional analysis in yeast verified that the two genes significantly contributed to the detoxification of glyphosate, and CsGSTU8-1 had a stronger role in detoxification than CsGSTU8-2. Taken together, these findings provide insights into the molecular responses of tea plants to glyphosate and the functions of CsGSTU8s in glyphosate detoxification, which can be used as a promising genetic resource for improving herbicide resistance in tea cultivars.

The Laccase Family Gene CsLAC37 Participates in Resistance to Colletotrichum gloeosporioides Infection in Tea Plants.

Fungal attacks have become a major obstacle in tea plantations. Colletotrichum gloeosporioides is one of the most devastating fungal pathogens in tea plantations that can severely affect tea yield and quality. However, the molecular mechanism of resistance genes involved in anthracnose is still largely unknown in tea plants. Here, we found that the laccase gene CsLAC37 was involved in the response to fungal infection based on a transcriptome analysis. The full-length CDS of CsLAC37 was cloned, and its protein sequence had the closest relationship with the Arabidopsis AtLAC15 protein compared to other AtLACs. Tissue-specific expression analysis showed that CsLAC37 had higher expression levels in mature leaves and stems than in the other tissues. Subcellular localization showed that the CsLAC37 protein was predominantly localized in the cell membrane. The expression levels of CsLAC37 were upregulated at different time points under cold, salt, SA, and ABA treatments. qRT-PCR confirmed that CsLAC37 responded to both Pestalotiopsis-like species and C. gloeosporioides infections. Functional validation showed that the hydrogen peroxide (H2O2) content increased significantly, and POD activity decreased in leaves after antisense oligonucleotide (AsODN) treatment compared to the controls. The results demonstrated that CsLAC37 may play an important role in resistance to anthracnose, and the findings provide a theoretical foundation for molecular breeding of tea varieties with resistance to fungal diseases.

Soil enzyme activities, physiological indicators, agronomic traits and yield of common buckwheat under herbicide combined with safeners.

In the pursuit of green agricultural development, alleviating the harmful effects of herbicides is critical. Herbicide safeners have been identified as an effective solution to safeguard crops without compromising the herbicidal efficacy. However, the impact of combined applications of herbicide and safeners on the physiological characteristics, growth, yield of common buckwheat, and soil enzyme activities remains unclear. Therefore, a two-year (2021 and 2022) field experiment was conducted in the Loess Plateau region of Northwest China under seven treatments: herbicide metolachlor application alone (H1); herbicide metolachlor combined with gibberellin (H1S1); herbicide metolachlor combined with brassinolide (H1S2); herbicide metolachlor combined with naian (H1S3); herbicide metolachlor combined with jiecaotong (H1S4); manual weeding (CK1) and spraying the same volume of water (CK2). The results indicated that H1S3 minimized herbicide toxicity while sustaining the herbicide control efficacy. H1S2 treatment significantly increased the chlorophyll content (SPAD value), superoxide dismutase (SOD), peroxidase (POD) activities, and decreased the malondialdehyde (MDA) content of the leaves compared to H1 treatment. Additionally, the safeners helped restore the biochemical homeostasis of the soil by preventing the inhibition of invertase and urease activities and increasing soil catalase activity. Furthermore, H1S2 promotion of dry matter accumulation, alleviation of herbicide inhibition on plant height, stem diameter, grainnumber per plant and thousand-grain weight resulted in a significant increase in grain yield (14.36 % in 2021 and 27.78 % in 2022) compared to other safener treatments. Overall, this study demonstrates that brassinolide as a safener can effectively mitigate the negative effects of herbicide on the growth and development of common buckwheat while also improving grain yield. These findings provide valuable technical guidance for sustainable and intensive production of common buckwheat in the Loess Plateau of Northwest China.

Effects of biochar coupled with chemical and organic fertilizer application on physicochemical properties and in vitro digestibility of common buckwheat (Fagopyrum esculentum Moench) starch.

Common buckwheat starch, a functional ingredient, has wide food and non-food applications. Excessive chemical fertilizer application during grain cultivation decreases quality. This study examined the effects of different combinations of chemical fertilizer, organic fertilizer, and biochar treatment on the physicochemical properties and in vitro digestibility of starch. The amendment of both organic fertilizer and biochar was observed to have a greater impact on the physicochemical properties and in vitro digestibility of common buckwheat starch in comparison to organic fertilizer amendment solely. The combined application of biochar, chemical, and organic nitrogen in an 80:10:10 ratio significantly increased the amylose content, light transmittance, solubility, resistant starch content, and swelling power of the starch. Simultaneously, the application reduced the proportion of amylopectin short chains. Additionally, this combination decreased the size of starch granules, weight-average molecular weight, polydispersity index, relative crystallinity, pasting temperature, and gelatinization enthalpy of the starch compared to the utilization of chemical fertilizer alone. The correlation between physicochemical properties and in vitro digestibility was analyzed. Four principal components were obtained, which accounted for 81.18 % of the total variance. These findings indicated that the combined application of chemical fertilizer, organic fertilizer, and biochar would improve common buckwheat grain quality.

Suitable nitrogen fertilizer application drives the endosperm development and starch synthesis to improve the physicochemical properties of common buckwheat grain.

The effects of nitrogen (N) fertilizer on endosperm development, starch component, key enzyme activity and grain quality of common buckwheat were investigated in this study. The results showed that N fertilization significantly enhanced the number and area of endosperm cells, and significant increases were also observed in the contents of amylose, amylopectin and total starch. With increasing N level, the activities of key enzyme significantly increased showing the maximum under the N2 level (180 kg N ha-1), and then decreased under high N level. As N level increased, the ash, crude protein and amylose content varied from 1.36 to 2.25 %, from 7.99 to 15.84 % and from 22.69 to 27.64 %, respectively. The gelatinization enthalpy significantly increased with the range of 3.46-5.66 J/g, while no change was found in crystalline structure of common buckwheat flour. These results indicated that appropriate N application could effectively improve the endosperm development, starch synthesis and accumulation, and grain properties of common buckwheat, with the best effect under the level of 180 kg N ha-1.

Relationship between nitrogen fertilizer and structural, pasting and rheological properties on common buckwheat starch.

Nitrogen is an essential element for the yield and quality of grain. In this study, the structural and physicochemical properties of two common buckwheat varieties under four nitrogen levels (0, 90, 180, 270 kg N ha-1) at one location in two years were investigated. With increasing nitrogen level, the contents of moisture and amylose decreased but the contents of ash and crude protein increased. Excessive nitrogen application significantly increased the granule size, but reduced the light transmittance, water solubility, swelling power, absorption of water and oil. All the samples showed a typical A - type pattern, while high relative crystallinity and low order degree were observed under high nitrogen level. The samples under high nitrogen level had lower textural properties, pasting properties and rheological properties but higher pasting temperature and gelatinization enthalpy. These results indicated that nitrogen fertilizer significantly affected the structural and physicochemical properties of common buckwheat starch.

Analysis of synthesis, accumulation and physicochemical properties of Tartary buckwheat starches affected by nitrogen fertilizer.

Nitrogen fertilizer is a crucial factor affecting the growth and grain quality of Tartary buckwheat. This study was to investigate the synthesis, accumulation, and physicochemical properties of Tartary buckwheat starches under four nitrogen levels (0, 90, 180, 270 kg N ha-1). The results showed that activities of four key enzymes, starch contents all first increased and then decreased with increasing nitrogen levels, and peaked at 180 kg N ha-1. All the starches showed typical A-type, while higher nitrogen levels significantly increased the relative crystallinity. The viscosities significantly decreased, onset, peak, and conclusion first decreased and then increased, while pasting temperature and gelatinization enthalpy increased with increasing nitrogen levels. Nitrogen fertilizer and year had significant effects on the synthesis, accumulation and physicochemical properties of Tartary buckwheat starch, and the nitrogen level of 180 kg N ha-1 was more suitable for planting in the northern area of the Loess Plateau.

Diverse effects of nitrogen fertilizer on the structural, pasting, and thermal properties of common buckwheat starch.

At present, the yield of common buckwheat, which is mainly grown in northern Shaanxi of China, is low and the grain quality is poor. Nitrogen is an important nutrient for the growth of common buckwheat, and appropriate nitrogen application can improve the grain quality. Nitrogen fertilizer could alter the starch granule morphology shapes and the granule size distribution. With increasing nitrogen levels, branch number, flower clusters number, grain number per plant, contents of protein and fat, size distribution of "C" granules, and percentages of light transmittance significantly increased, whereas amylose content and retrogradation decreased. All the samples displayed typical A-type X-ray diffraction patterns. Starch showed higher pasting temperature and gelatinization enthalpy but lower trough and final viscosities under high nitrogen levels. These results suggested N2 treatment was more suitable for common buckwheat growth, principal components and correlation analysis revealed that nitrogen fertilizer significantly affected the physicochemical properties of common buckwheat starches.

Soil properties, bacterial and fungal community compositions and the key factors after 5-year continuous monocropping of three minor crops.

Minor grain crops are widely cultivated in northwest China and played important roles in local economic. Soil microbes play a central role in ecological function and biological stability and related to soil quality. In order to uncover the soil microbial composition differences and the factors under 5-year continuous monocropping of three minor crops (Proso millet, Common bean and Common buckwheat) in Guan-Zhong Plain, six soil nutrimental parameters, soil pH, soil moisture content, and four soil enzyme activities were analyzed and soil microbial composition were sequenced. The results showed that after 5-years of continuous monocropping, different cover crops influenced most of soil physicochemical properties, expect soil moisture content (P < 0.05), the available nutrients were significant higher in proso millet soil, and the pH was significantly higher in common buckwheat soil. soil ALP, catalase and urease activities were significantly different between soils (P< 0.01), in which soil catalase activities were significantly lower and soil ALP and urease activities were significantly higher than that of proso millet and common buckwheat. A total of 171439 sequences, 9468 OTUs and 29 phylum for bacteria, 128920 sequences, 544 OTUs and 27 phylum for fungi were obtained. In addition, no significantly difference obtained in diversity and richness between soils (P < 0.05). According to relative abundance, Proteobacteria, Chloroflexi, Gemmatimonadetes and Acidobacteria were the dominant bacterial phylum in all samples, moreover, the relative abundance of Caldiserica was significantly different between soils (P < 0.05). Ascomycota (79.04%-90.21%) was dominant phylum in fungal community and phylum Phragmoplastophyta (P < 0.01) and Glomeromycota (P < 0.05) were significantly different between soils. Redundancy analysis indicated that available nutrients Nitrogen and Potassium are the strongest predictors in both bacterial and fungal community. In conclusion, different cover crops influenced soil nutrient properties, soil pH and soil microbial composition, and continuous monocropping decreased soil fertility condition. Moreover, Common bean and Common buckwheat were more sensitive to monocropping treatment.

[Control effects of different herbicides on weeds as well as their effects on growth and deve-lopment of broomcorn millet]. / 不同除草剂的田间杂草防效及对糜子生长发育的影响.

Selecting a herbicide suitable for broomcorn millet is a key issue in high efficiency and safe production of broomcorn millet and ecological environment improvement. We compared the control effects of 22 different herbicides on weeds as wells as their effects on growth and development of glutinous millet (Yumi 2) in the field. The results showed that, 1) soil applied Goyou, Butachlor, atrazine, and Benzipram, and stem- and leaf-applied WP mixture of tribenuron-methyl·carfentrazone-ethyl·MCPA-Na, Sigma Broad, Tengjing, Taoshi·Youxian and Kuofei had little phytotoxi-city, with broomcorn millet seedlings growing normally, and the other herbicides had phytotoxicities on broomcorn millet. 2) All the herbicides controlled weeds in the field to some extent, with better performance of soil applied herbicides than the stem- and leaf-applied ones, and affected plant height, functional leaf SPAD and spike weight per plant of broomcorn millet. 3) Compared with manual weeding, all herbicides reduced yields of glutinous millet. Compared with no herbicides application, some herbicides had yield-increasing effects. Among the soil applied herbicides, Go-you, Butachlor, atrazine, and Benzipram performed better in weeding control, increasing the yield of broomcorn millet by more than 60% compared with no herbicide control. Among the stem- and leaf-applied herbicides, Sigma Broad and WP mixture of tribenuron-methyl·carfentrazone-ethyl·MCPA-Na performed better in weeding control, increasing the yield of glutinous millet by more than 50% compared with no herbicide control. Therefore, applying 38% atrazine or 44% Monosulfuron to soil before the emergence of broomcorn millet, or stem- and leaf-applied 3.6% mesosulfuron-methyl or 55% WP mixture of tribenuron-methyl·carfentrazone-ethyl·MCPA-Na after the emergence of broomcorn millet, performed better in weeding control and with limited effects on the growth and development of broomcorn millet.

Structural, pasting and thermal properties of common buckwheat (Fagopyrum esculentum Moench) starches affected by molecular structure.

Common buckwheat starch (CBS) has extensive using value in the human diet. In this study, the molecular structure and physicochemical properties of CBS isolated from five cultivars collected from three regions of China were studied. Variations in molecular structure, crystalline structure, complexity, water solubility (WS), swelling power (SP), pasting properties, and thermal characteristics were recorded among the starches. The CBS had both similarities and differences in its properties by comparison with maize starch (MS) and potato starch (PS). The average molecular weight (MW) and amylopectin average chain length (ACL) of CBS ranged from 3.86 × 107 g/mol to 4.68 × 107 g/mol and from 21.29% to 22.68%, respectively. CBS and MS were divided into one subgroup and showed typical A diffraction patterns, while PS was divided into two subgroups and exhibited a typical B polymorphic pattern. The WS and SP of all the starches significantly increased with increasing temperature and had great variation at 70 °C and 90 °C. Pearson's correlation analysis showed that the molecular structure of starches greatly affected the physicochemical properties. This study revealed that the physicochemical properties of CBS could be affected by the molecular structures.

Common buckwheat-resistant starch as a suitable raw material for food production: A structural and physicochemical investigation.

Heat-moisture treatment (HMT) of starch is defined as a physical method to change its properties. Compared with maize and potato, starches from common buckwheat (Xinong9976 and Pingqiao2) were isolated and its morphology and physicochemical properties investigated by scanning electron microscope (SEM), X-ray diffraction (XRD), ATR-FTIR analysis, rapid viscosity analyzer (RVA) and differential scanning calorimeter (DSC) were studied before and after HMT. The experimental results showed that there were obvious differences between native starch (NS) and resistant starch (RS) of common buckwheat. HMT altered the A-type crystalline pattern and the degree of short-range order of common buckwheat starches and significantly decreased water solubility, swelling power (70-90 °C), freeze-thaw stability and pasting properties and increased oil and water absorption capacities, light transmittance as well as thermal stability. This study shows that the NS and RS of common buckwheat can be used as the suitable raw materials in food processing.

Effects of phosphate fertiliser on the physicochemical properties of Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) starch.

Phosphate fertilisation affects the growth, development and quality of Tartary buckwheat. In this study, the effect of different phosphorus levels, including 0, 15, 75, and 135 kg/ha (non-, low-, medium-, and high-phosphorus levels, respectively), on the characteristics of starch from Tartary buckwheat were investigated in 2015 and 2017. With increased phosphorus level, the median diameter of starch granules and the apparent amylose content initially decreased and then increased. All starch samples showed the features of A-type X-ray diffraction patterns. Starches under medium-phosphorus treatment showed higher relative crystallinity than those under non-phosphorus treatment, as well as the highest solubility, gelatinisation enthalpy and transmittance among all starches. Starches under low-phosphorus treatment exhibited higher pasting properties than those under non-phosphorus treatment. This research revealed that phosphorus treatments and year significantly affected the physicochemical properties of Tartary buckwheat starch, and can provide information for the applications of starch in the food and non-food industries.

Identification of Differentially Expressed Genes Involved in the Molecular Mechanism of Pericarp Elongation and Differences in Sucrose and Starch Accumulation between Vegetable and Grain Pea (Pisum sativum L.).

Pea (Pisum sativum L.), as a major source of plant protein, is becoming one of the major cultivated crop species worldwide. In pea, the pericarp is an important determinant of the morphological characteristics and seed yield. To investigate the molecular mechanism of pericarp elongation as well as sucrose and starch accumulation in the pods of different pea cultivars, we performed transcriptomic analysis of the pericarp of two types of pea cultivar (vegetable pea and grain pea) using RNA-seq. A total of 239.44 Gb of clean sequence data were generated, and were aligned to the reference genome of Pisum sativum L. In the two samples, 1935 differentially expressed genes (DEGs) were identified. Among these DEGs, three antioxidant enzyme superoxide dismutase (SOD) were detected to have higher expression levels in the grain pea pericarps at the pod-elongating stages. Otherwise, five peroxidase (POD)-encoding genes were detected to have lower expression levels in the vegetative pericarps at the development stage of pea pod growth. Furthermore, genes related to starch and sucrose metabolism in the pea pod, such as SUS, INV, FBA, TPI, ADPase, SBE, SSS, and GBSS, were found to be differentially expressed. The RNA-seq data were validated through real-time quantitative RT-PCR of 13 randomly selected genes. Our findings provide the gene expression profile of, as well as differential expression information on, the two pea cultivars, which will lay the foundation for further studies on pod development and nutrition accumulation in the pea and provide valuable information for pea cultivar improvement.

Comparative analysis of proso millet (Panicum miliaceum L.) leaf transcriptomes for insight into drought tolerance mechanisms.

BACKGROUND: Drought stress is a major abiotic stress that causes huge losses in agricultural production. Proso millet (Panicum miliaceum L.) can efficiently adapt to drought stress and provides important information and gene resources to improve drought tolerance. However, its complex drought-responsive mechanisms remain unclear. RESULTS: Among 37 core Chinese proso millet cultivars, Jinshu 6 (JS6) was selected as the drought-sensitive test material, whereas Neimi 5 (NM5) was selected as the drought-tolerant test material under PEG-induced water stress. After sequencing, 1695 differentially expressed genes (DEGs) were observed in JS6 and NM5 without PEG-induced water stress (JS6CK and NM5CK). A total of 833 and 2166 DEGs were found in the two cultivars under simulated drought by using 20% PEG-6000 for 6 (JS6T6 and NM5T6) and 24 h (JS6T24 and NM5T24), respectively. The DEGs in JS6T6 and JS6T24 treatments were approximately 0.298- and 0.754-fold higher than those in NM5T6 and NM5T24, respectively. Compared with the respective controls, more DEGs were found in T6 treatments than in T24 treatments. A delay in the transcriptional responses of the ROS scavenging system to simulated drought treatment and relatively easy recovery of the expression of photosynthesis-associated genes were observed in NM5. Compared with JS6, different regulation strategies were observed in the jasmonic acid (JA) signal transduction pathway of NM5. CONCLUSION: Under PEG-induced water stress, NM5 maintained highly stable gene expression levels. Compared with drought-sensitive cultivars, the different regulation strategies in the JA signal transduction pathway in drought-tolerant cultivars may be one of the driving forces underlying drought stress tolerance.

Effects of nitrogen level on the physicochemical properties of Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) starch.

Nitrogen is an essential nutrient for Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) because this substance affects the yield and quality of Tartary buckwheat. The physicochemical characteristics of starch represent the important parameters of Tartary buckwheat quality. The effects of different levels of nitrogen application at different levels (0, 45, 135 and 225kg/ha in 2015 and 0, 90, 180 and 270kg/ha in 2017) on the physicochemical characteristics of Tartary buckwheat starch were studied. The amylose content, particle size and retrogradation of Tartary buckwheat starch with nitrogen were lower than those of without nitrogen. The structure complexity, pasting temperature, gelatinization enthalpy, relative crystallinity, light transmittance and solubility of the former were higher than those of the latter. Nitrogen application did not change the 'A'-type crystalline pattern of Tartary buckwheat starch. This study indicated that nitrogen level and years and the interactions among nitrogen fertilizer levels and years significantly affected the physicochemical properties of Tartary buckwheat starch. These integrated results also provided information about the management of fertilization conditions to obtain starches with special properties for applications in food or nonfood industries.

Tartary buckwheat (Fagopyrum tataricum Gaertn.) starch, a side product in functional food production, as a potential source of retrograded starch.

A starch rich fraction is a side product in Tartary buckwheat processing. This study investigated the fractions that are of technological and nutritional interest. Tartary buckwheat starch granules had a diameter of 3-14 µm, and presented a typical type "A" X-ray diffraction pattern. They contained nearly 39.0% amylose. The solubility of Tartary buckwheat starch was much lower at 70-90 °C (ranging within 9.9-10.4% at 90 °C) than that in maize (up to 49.3%) and potato (up to 85.0%) starch. The starch of one variety of Tartary buckwheat had significantly lower solubility at 70 °C and 80 °C than that of common buckwheat. The starch peak viscosity and breakdown were higher and pasting time was shorter in Tartary buckwheat than in that of the starch of common buckwheat. Tartary buckwheat starch had unique pasting and physicochemical properties, and is thereby capable of being exploited as a suitable raw material of retrograded starch in food processing.

[Border effect and physiological characteristics of broomcorn millet under film mulching on ridge-furrow for harvesting rainwater model in the semi-arid region of Northern Shaanxi, China].

To explore the border effect and physiological characteristic of broomcorn millet growing under different film mulching on ridge-furrow for harvesting rainwater models in the semi-arid region of Northern Shaanxi, China, a three-year field experiment was conducted with four different widths of ridge and furrow, and the bare land flat sowing as the control (NM). The width of ridge and furrow varied as ridge: furrow = 40 cm: 40 cm (P40), 60 cm: 60 cm (P60), 80 cm: 80 cm (P80), and 100 cm:100 cm (P100). The results showed that the wider the width of furrow and ridge was, the stronger the border advantage and the border effect index of the yield were. With the increase in width of furrow and ridge, the yield increasing effect of side rows increased with the maximum of 207.7%, and the yield increasing effect of middle rows decreased with the minimum of 10.3%. P60 reached the highest yield within three years. The yield contribution rate of side rows was higher than that of middle rows (P < 0.05). The chlorophyll contents, Ch1 a/Ch1 b, and photosynthetic rate of side rows were higher than those of middle rows among the different harvesting rainwater models. The wider the width of furrow and ridge was, the stronger the photosynthetic capacity of side rows was, and the weaker the photosynthetic capacity of middle rows was. The optimal type of ridge and furrow was P60 in the semi-arid region of Northern Shaanxi.