Metabolomic analyses reveal substances that contribute to the increased freezing tolerance of alfalfa (Medicago sativa L.) after continuous water deficit.

BACKGROUND: Alfalfa is a high-quality forage cultivated widely in northern China. Recently, the failure of alfalfa plants to survive the winter has caused substantial economic losses. Water management has attracted considerable attention as a method for the potential improvement of winter survival. The aim of this study was to determine whether and how changes in the water regime affect the freezing tolerance of alfalfa. RESULTS: The alfalfa variety WL353LH was cultivated under water regimes of 80 and 25% of water-holding capacity, and all the plants were subjected to low temperatures at 4/0 °C (light/dark) and then - 2/- 6 °C (light/dark). The semi-lethal temperatures were lower for water-stressed than well-watered alfalfa. The pool sizes of total soluble sugars, total amino acids, and proline changed substantially under water-deficit and low-temperature conditions. Metabolomics analyses revealed 72 subclasses of differential metabolites, among which lipid and lipid-like molecules (e.g., fatty acids, unsaturated fatty acids, and glycerophospholipids) and amino acids, peptides, and analogues (e.g., proline betaine) were upregulated under water-deficit conditions. Some carbohydrates (e.g., D-maltose and raffinose) and flavonoids were also upregulated at low temperatures. Finally, Kyoto Encyclopedia of Genes and Genomes analyses revealed 18 significantly enriched pathways involved in the biosynthesis and metabolism of carbohydrates, unsaturated fatty acids, amino acids, and glycerophospholipids. CONCLUSIONS: Water deficit significantly enhanced the alfalfa' freezing tolerance, and this was correlated with increased soluble sugar, amino acid, and lipid and lipid-like molecule contents. These substances are involved in osmotic regulation, cryoprotection, and the synthesis, fluidity, and stability of the cellular membrane. Our study provides a reference for improving alfalfa' winter survival through water management.

SERK genes identification and expression analysis during somatic embryogenesis and sporogenesis of sexual and apomictic Brachiaria brizantha (Syn. Urochloa brizantha).

MAIN CONCLUSION: In Brachiaria brizantha BbrizSERK1, BbrizSERK2 and BbrizSERK3 were identified. SERK expression marks somatic embryogenesis, sexual MMC, and sexual and apomictic PMC. BbrizSERK3 might have a regulatory role in reproductive development. Somatic embryogenesis receptor-like kinase (SERK) consists of plasma membrane receptor genes that have been characterized in various species, associated with several aspects of plant development, including reproduction. SERK genes are involved in anther development and in early embryo development in sexual and asexual seed formation. To comprehend the complexity of the SERK genes and their function in Brachiaria reproduction, we performed a homology-based search in a genomic database of a sexual B. brizantha and identified sequences of three SERK genes, BbrizSERK1, BbrizSERK2, and BbrizSERK3. RNASeq data showed equivalent abundance of BbrizSERK1 and BbrizSERK2 transcripts in ovaries at early megasporogenesis of sexuals and apomicts, while BbrizSERK3 transcripts were more abundant in ovaries of sexuals than in apomicts. BbrizSERK3 results in three coding sequences due to alternative splicing, among them Variant 1 results in a protein with all the predicted domains of a SERK. BbrizSERK transcripts were detected in male reproductive tissues of both sexual and apomictic plants, suggesting a role in controlling anther development. BbrizSERK transcripts were detected early in ovule development, in the integuments, and in the megaspore mother cell of the sexual plant, but not in the cells that give rise to apomictic embryo sacs, suggesting a role in female reproductive development of sexuals. This paper provides evidences that SERK genes plays a role in the onset and establishment of somatic embryogenesis and in the reproductive development of B. brizantha and suggests a distinct role of BbrizSERK in apomixis initiation.

Exogenous jasmonic acid decreased Cu accumulation by alfalfa and improved its photosynthetic pigments and antioxidant system.

Jasmonic acid (JA) is an important phytohormone, which among others may be involved in the regulation of plant accumulating heavy metal. This experiment was designed to explore the effects of exogenous JA on the responses of alfalfa to Cu stress (100 µM) in Hoagland solution. When 1, 5 or 10 mM JA was added to the treatment with Cu addition, Cu concentrations in roots and leaves of alfalfa were significantly decreased (p < 0.05) to some extents compared to the treatment without JA addition. The biomasses of roots and leaves of alfalfa in treatments of JA additions were significantly increased (p < 0.05) compared to the Cu stress treatment. Similarly, the concentrations of Chlorophyll, antioxidant enzyme activities, MDA and H2O2 were improved accordingly. But these factors of JA were not improved further when its concentration added in media was the highest (10 mM), indicating its improvement roles were limited. These results suggested that there were positive roles of exogenous JA on alfalfa decreased its Cu accumulation and toxicities might via reduced oxidative stress.

Improved feeding and forages at a crossroads: Farming systems approaches for sustainable livestock development in East Africa.

Dairy development provides substantial potential economic opportunities for smallholder farmers in East Africa, but productivity is constrained by the scarcity of quantity and quality feed. Ruminant livestock production is also associated with negative environmental impacts, including greenhouse gas (GHG) emissions, air pollution, high water consumption, land-use change, and loss of biodiversity. Improved livestock feeding and forages have been highlighted as key entry point to sustainable intensification, increasing food security, and decreasing environmental trade-offs including GHG emission intensities. In this perspective article, we argue that farming systems approaches are essential to understand the multiple roles and impacts of forages in smallholder livelihoods. First, we outline the unique position of forages in crop-livestock systems and systemic obstacles to adoption that call for multidisciplinary thinking. Second, we discuss the importance of matching forage technologies with agroecological and socioeconomic contexts and niches, and systems agronomy that is required. Third, we demonstrate the usefulness of farming systems modeling to estimate multidimensional impacts of forages and for reducing agro-environmental trade-offs. We conclude that improved forages in East Africa are at a crossroads: if adopted by farmers at scale, they can be a cornerstone of pathways toward sustainable livestock systems in East Africa.

Root-zone hypoxia reduces growth of the tropical forage grass Urochloa humidicola in high-nutrient but not low-nutrient conditions.

BACKGROUND AND AIMS: The perennial C4 grass Urochloa humidicola is widely planted on infertile acidic and waterlogging-prone soils of tropical America. Waterlogging results in soil anoxia, and O2 deficiency can reduce nutrient uptake by roots. Interestingly, both nutrient deficiencies and soil waterlogging can enhance root cortical cell senescence, and the increased gas-filled porosity facilitates internal aeration of roots. We tested the influence of nutrient supply and root-zone O2 on root traits, leaf nutrient concentrations and growth of U. humidicola. METHODS: Plants were grown in pots in a completely randomized design under aerated or stagnant deoxygenated hydroponic conditions and six nutrient regimes, with low to high concentrations of all essential elements, for 28 d in a controlled-temperature greenhouse. The standard acid solution (SAS) used was previously designed based on infertile acidic soils of the tropical America savannas, and step increases in the concentration of SAS were used in aerated or deoxygenated 0.1 % agar solution, which mimics changes in gas composition in waterlogged soils. Measurements included shoot and root growth, root porosity, root anatomy, radial O2 loss, and leaf tissue nutrient concentrations. KEY RESULTS: Shoot dry mass was reduced for plants in stagnant compared with aerated conditions at high, but not at low, levels of mineral nutrition. In low-nutrition stagnant solution, roots were shorter, of greater porosity and had smaller radial thickness of the stele. Suberized lamellae and lignified sclerenchyma, as well as a strong barrier to radial O2 loss, were documented for roots from all treatments. Leaf nutrient concentrations of K, Mg and Ca (but not N, P and S) were higher in aerated than in stagnant conditions. CONCLUSIONS: Under low-nutrient conditions, plant growth in stagnant solution was equal to that in aerated solution, whereas under higher-nutrient regimes growth increased but dry mass in stagnant solution was less than in aerated solution. Slow growth in low-nutrient conditions limited any further response to the low O2 treatment, and greater porosity and smaller stele size in roots would enhance internal O2 movement within roots in the nutrient-limited stagnant conditions. A constitutive barrier to radial O2 loss and aerenchyma facilitates O2 movement to the tips of roots, which presumably contributes to maintaining nutrient uptake and the tolerance of U. humidicola to low O2 in the root-zone.

Differential transferability of EST-SSR primers developed from the diploid species Pseudoroegneria spicata, Thinopyrum bessarabicum, and Thinopyrum elongatum.

Simple sequence repeat technology based on expressed sequence tag (EST-SSR) is a useful genomic tool for genome mapping, characterizing plant species relationships, elucidating genome evolution, and tracing genes on alien chromosome segments. EST-SSR primers developed from three perennial diploid species of Triticeae, Pseudoroegneria spicata (Pursh) Á. Löve (having St genome), Thinopyrum bessarabicum (Savul. & Rayss) Á. Löve (Jb = Eb = J), and Thinopyrum elongatum (Host) D.R. Dewey (Je = Ee = E), were used to produce amplicons in these three species to (i) assess relative transferability, (ii) identify polymorphic species-specific markers, and (iii) determine genome relationships among the three species. Because of the close relationship between Jb and Je genomes, EST-SSR primers derived from Th. bessarabicum and Th. elongatum had greater transferability to each other than those derived from the St-genome P. spicata. A large number of polymorphic species- and genome-specific EST-SSR amplicons were identified that will be used for construction of genetic maps of these diploid species, and tracing economically useful genes in breeding or gene transfer programs in various species of Triticeae.

Dynamics of Neotyphodium uncinatum and N-formylloline in Italian ryegrass, and their relation to insect resistance in the field.

AIMS: A fungal endophyte, Neotyphodium uncinatum, accumulates N-formylloline, which is toxic to Hemipteran insects, in Italian ryegrass. This study aimed to clarify the dynamics of N. uncinatum and N-formylloline in Italian ryegrass, and their relationship to insect resistance. METHODS AND RESULTS: Changes in the density and localization of N. uncinatum and N-formylloline in N. uncinatum-infected Italian ryegrass were examined by real-time PCR and gas chromatography, respectively. Neotyphodium uncinatum multiplied on pseudostems at the flowering stage, and then increased on inflorescences at the ripening stage. On the other hand, N-formylloline accumulated heavily in inflorescences and leaf blades, but lightly in pseudostems at the ripening stage. In field experiments, N. uncinatum-infected Italian ryegrass suppressed the occurrence of Stenotus rubrovittatus, which fed on the inflorescences, but was not effective to Laodelphax striatellus, which do not necessarily prefer a particular plant tissue. CONCLUSION: Localization of N. uncinatum and N-formylloline were discordant in Italian ryegrass. The N. uncinatum-infected Italian ryegrass was effective to only insects that prefer to feed on particular plant tissues containing N-formylloline. SIGNIFICANCE AND IMPACT OF STUDY: Our data implies that the relationship between insect habits and the dynamics of alkaloids in plants is important for the effective use of endophyte-infected crops.

Green control for inhibiting Rhizopus oryzae growth by stress factors in forage grass factory.

The forage grass factory could break through the restrictions of land resources, region and climate to achieve efficient production throughout the year by accurate and intelligent management. However, due to its closed environment, mold outbreaks in the forage grass factory were severe, significantly affecting barley production. In this study, 9 contaminated barley tissues were collected and 45 strains were isolated in forage grass factory. After ITS sequencing, 45 strains were all identified as Rhizopus oryzae. Through stress factor assays, R. oryzae growth was seriously hindered by low concentration of sodium nitrate, high pH value and ozone water treatment. High pH and ozone water affected growth mainly by altering membrane integrity of R. oryzae. Sodium nitrate inhibited the growth of R. oryzae mainly by affecting the amount of sporulation. Low concentration of sodium nitrate and ozone water did not affect the growth of barley. High concentrations of sodium nitrate (100 mM) and pH values (8-8.5) inhibited barley growth. Among them, ozone water had the most obvious inhibition effect on R. oryzae. Large-scale ozone water treatment in the forage grass factory had also played a role in restoring barley production. Taken together, the green techonology to control mold disease and maintain the safety of forage through different physicochemical methods was selected, which was of considerable application value in animal husbandry.

Response to Waterlogging Stress in Wild and Domesticated Accessions of Timothy (Phleum pratense) and Its Relatives P. alpinum and P. nodosum.

Timothy (Phleum pratense) is a cool-season perennial forage grass widely grown for silage and hay production in northern regions. Climate change scenarios predict an increase in extreme weather events with fluctuating periods of high rainfall, requiring new varieties adapted to waterlogging (WL). Wild accessions could serve as germplasm for breeding, and we evaluated the responses of 11 wild and 8 domesticated accessions of timothy, P. nodosum and P. alpinum from different locations in northern Europe. Young plants at tillering stage were exposed to WL for 21 days in a greenhouse, and responses in growth allocation and root anatomy were studied. All accessions produced adventitious roots and changed allocation of growth between shoot and root as a response to WL, but the magnitude of these responses varied among species and among accessions. P. pratense responded less in these traits in response to WL than the other two species. The ability to form aerenchyma in the root cortex in response to WL was found for all species and also varied among species and among accessions, with the highest induction in P. pratense. Interestingly, some accessions were able to maintain and even increase root growth, producing more leaves and tillers, while others showed a reduction in the root system. Shoot dry weight (SDW) was not significantly affected by WL, but some accessions showed different and significant responses in the rate of production of leaves and tillers. Overall correlations between SDW and aerenchyma and between SDW and adventitious root formation were found. This study identified two wild timothy accessions and one wild P. nodosum accession based on shoot and root system growth, aerenchyma formation and having a root anatomy considered to be favorable for WL tolerance. These accessions are interesting genetic resources and candidates for development of climate-resilient timothy varieties.

Chromosome-level reference genome assembly provides insights into the evolution of Pennisetum alopecuroides.

Pennisetum alopecuroides is an important forage grass resource, which plays a vital role in ecological environment improvement. Therefore, the acquisition of P. alopecuroides genome resources is conducive to the study of the adaptability of Pennisetum species in ecological remediation and forage breeding development. Here we assembled a P. alopecuroides cv. 'Liqiu' genome at the chromosome level with a size of approximately 845.71 Mb, contig N50 of 84.83Mb, and genome integrity of 99.13% as assessed by CEGMA. A total of 833.41-Mb sequences were mounted on nine chromosomes by Hi-C technology. In total, 60.66% of the repetitive sequences and 34,312 genes were predicted. The genomic evolution analysis showed that P. alopecuroides cv. 'Liqiu' was isolated from Setaria 7.53-13.80 million years ago and from Cenchrus 5.33-8.99 million years ago, respectively. The whole-genome event analysis showed that P. alopecuroides cv. 'Liqiu' underwent two whole-genome duplication (WGD) events in the evolution process, and the duplication events occurred at a similar time to that of Oryza sativa and Setaria viridis. The completion of the genome sequencing of P. alopecuroides cv. 'Liqiu' provides data support for mining high-quality genetic resources of P. alopecuroides and provides a theoretical basis for the origin and evolutionary characteristics of Pennisetum.

Improving abiotic stress tolerance of forage grasses - prospects of using genome editing.

Due to an increase in the consumption of food, feed, and fuel and to meet global food security needs for the rapidly growing human population, there is a necessity to obtain high-yielding crops that can adapt to future climate changes. Currently, the main feed source used for ruminant livestock production is forage grasses. In temperate climate zones, perennial grasses grown for feed are widely distributed and tend to suffer under unfavorable environmental conditions. Genome editing has been shown to be an effective tool for the development of abiotic stress-resistant plants. The highly versatile CRISPR-Cas system enables increasingly complex modifications in genomes while maintaining precision and low off-target frequency mutations. In this review, we provide an overview of forage grass species that have been subjected to genome editing. We offer a perspective view on the generation of plants resilient to abiotic stresses. Due to the broad factors contributing to these stresses the review focuses on drought, salt, heat, and cold stresses. The application of new genomic techniques (e.g., CRISPR-Cas) allows addressing several challenges caused by climate change and abiotic stresses for developing forage grass cultivars with improved adaptation to the future climatic conditions. Genome editing will contribute towards developing safe and sustainable food systems.

Combined forage grass-microbial for remediation of strontium-contaminated soil.

Enrichment plants were screened from six forage grasses in this study to establish a complete combined forage grass-microbial remediation system of strontium-contaminated soil, and microbial groups were added to the screened dominant forage grasses. The occurrence states of strontium in forage grasses were explored by the BCR sequential extraction method. The results showed that the annual removal rate of Sudan grass (Sorghum sudanense (Piper) Stapf.) reached 23.05% in soil with a strontium concentration of 500 mg·kg-1. Three dominant microbial groups: E, G and H, have shown good facilitation effects in co-remediation with Sudan grass and Gaodan grass (Sorghum bicolor × sudanense), respectively. When compared to the control, the strontium accumulation of forage grasses in kg of soil with microbial groups was increased by 0.5-4 fold. The optimal forage grass-microbial combination can theoretically repair contaminated soil in three years. The microbial group E was found to promote the transfer of the exchangeable state and the reducible state of strontium to the overground part of the forage grass. Metagenomic sequencing results showed that the addition of microbial groups increased Bacillus spp. in rhizosphere soil, enhanced the disease resistance and tolerance of forage grasses, and improved the remediation ability of forage grass-microbial combinations.

Leymus chinensis resists degraded soil stress by modulating root exudate components to attract beneficial microorganisms.

Phytoremediation is an effective means to improve degraded soil nutrients and soil structure. Here, we investigated the remediation effects of Leymus chinensis on the physicochemical properties and structure of degraded soil after 3 years of cultivation and explored the bacterial and fungal drivers in root exudates by metabolomics and high-throughput sequencing. The results showed that root exudates increased soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP) and soil aggregates, and organic acids in root exudates reduced pH and activated insoluble nutrients into forms that are available to plants, such as available nitrogen (NH4 +-N), nitrate nitrogen (NO3 --N), and available phosphorus (AP). The cultivation of L. chinensis restored the diversity and richness of soil microorganisms and recruited potential beneficial bacteria and fungi to resist degraded soil stress, and L. chinensis also regulated the abundances of organic acids, amino acids and fatty acids in root exudates to remediate degraded soils. Spearman correlation analysis indicated that glutaric acid, 3-hydroxybutyric acid and 4-methylcatechol in root exudates attracted Haliangium, Nitrospira and Mortierella to the rhizosphere and dispersed the relative abundance of the harmful microorganisms Fusicolla and Fusarium. Our results demonstrate that L. chinensis enhances soil fertility, improves soil structure, promotes microbial diversity and abundance, and recruits potentially beneficial microorganisms by modulating root exudate components.

EST-SSR Primer Development and Genetic Structure Analysis of Psathyrostachys juncea Nevski.

Psathyrostachys juncea is a perennial forage grass which plays an important role in soil and water conservation and ecological maintenance in cold and dry areas of temperate regions. In P. juncea, a variety of biotic and abiotic stress related genes have been used in crop improvement, indicating its agronomic, economic, forage, and breeding value. To date, there have been few studies on the genetic structure of P. juncea. Here, the genetic diversity and population structure of P. juncea were analyzed by EST-SSR molecular markers to evaluate the genetic differentiation related to tillering traits in P. juncea germplasm resources. The results showed that 400 simple sequence repeat (SSR) loci were detected in 2,020 differentially expressed tillering related genes. A total of 344 scored bands were amplified using 103 primer pairs, out of which 308 (89.53%) were polymorphic. The Nei's gene diversity of 480 individuals was between 0.092 and 0.449, and the genetic similarity coefficient was between 0.5008 and 0.9111, with an average of 0.6618. Analysis of molecular variance analysis showed that 93% of the variance was due to differences within the population, and the remaining 7% was due to differences among populations. Psathyrostachys juncea materials were clustered into five groups based on population genetic structure, principal coordinate analysis and unweighted pair-group method with arithmetic means (UPGMA) analysis. The results were similar between clustering methods, but a few individual plants were distributed differently by the three models. The clustering results, gene diversity and genetic similarity coefficients showed that the overall genetic relationship of P. juncea individuals was relatively close. A Mantel test, UPGMA and structural analysis also showed a significant correlation between genetic relationship and geographical distribution. These results provide references for future breeding programs, genetic improvement and core germplasm collection of P. juncea.

Abscisic acid-polyacrylamide (ABA-PAM) treatment enhances forage grass growth and soil microbial diversity under drought stress.

Drought restricts the growth of alpine grassland vegetation. This study aimed to explore a new technical system to improve the drought resistance of forage grass. Qinghai cold-land Poa pratensis seedlings were used in the drought stress experiment. A combination of abscisic acid (ABA) and polyacrylamide (PAM) were used to affect the growth, leaf physiology, soil enzyme activity, and rhizosphere microbial diversity of P. pratensis. The fresh leaf weight and root surface area were significantly increased after ABA-PAM combined treatment, while root length was significantly reduced. Besides, the leaf catalase (CAT) and superoxide dismutase (SOD) enzyme activity, proline and chlorophyll content, increased after the treatment, while malondialdehyde (MDA) content decreased. The treatment also increased sucrase, urease, and alkaline protease activities in rhizosphere soil, while decreasing acid phosphatase and neutral phosphatase enzyme activities. ABA-PAM combined treatment enhanced the rhizosphere microbial community and forage drought resistance by altering the abundance of various dominant microorganisms in the rhizosphere soil. The relative abundances of Actinobacteria, Chloroflexi, and Acidobacteria decreased, while Proteobacteria, Firmicutes, and Ascomycota increased. Unlike the relative abundance of Gibberella that decreased significantly, Komagataeibacter, Lactobacillus, Pichia, and Dekkera were significantly increased. Single-factor collinearity network analysis revealed a close relationship between the different rhizosphere microbial communities of forage grass, after ABA-PAM treatment. This study implies that ABA-PAM combined treatment can improve the drought resistance of forages. Therefore, it provides a theoretical and practical basis for restoring drought-induced grassland degradation.

High Recovery from Either Waterlogging or Drought Overrides Any Beneficial Acclimation of Chloris gayana Facing a Subsequent Round of Stress.

Climate models predict that plants will face extreme fluctuations in water availability in future global change scenarios. Then, forage production will be more frequently subjected to the destabilizing pressure of sequentially occurring waterlogging and drought events. While the isolated effects of drought (D) and waterlogging (WL) are well characterized, little is known about the consequences when both stresses occur sequentially. We hypothesized that plants sequentially subjected to opposite water scenarios (D followed by WL or vice versa) are less stress tolerant than plants experiencing repetitions of the same type of water stress (i.e., D + D or WL + WL) due to contrasting acclimation and allocation to either shoots (WL) or roots (D). Chloris gayana (a tropical forage grass capable of tolerating either D and WL) plants were randomly assigned to nine treatments (a sequence of two stress rounds-WL or D-each followed by a recovery phase at field capacity). Relative growth rates and allometric responses were measured after each stress round and recovery period. In the first round of stress, both WL and D reduced plant RGR similarly, despite their allocation being opposite-prioritizing shoots or roots under WL and D, respectively. The high recovery displayed after either WL or D overrode any possible acclimation of the plants facing a second round of water stress. We conclude that the tolerance of C. gayana to sequential water stress (either for WL or D) is likely to depend more heavily on its recovery ability than on its previous adjustment to any stress scenario that may evoke memory responses. Knowledge like this could help improve forage grass breeding and the selection of cultivars for poorly drained soils subject to sequential stress events.

Study of the occurrence of toxic alkaloids in forage grass by liquid chromatography tandem mass spectrometry.

The toxic alkaloids in forage grass present a serious health hazard to humans and livestock, especially ergot alkaloids (EAs), pyrrolizidine alkaloids (PAs) and tropane alkaloids (TAs). Hence, there is a need for a simultaneous method that allows these dangerous plant toxins to be determined. A simple and effective method was developed to determine fifteen toxic alkaloids (EAs, PAs and TAs) in forage grass using the QuEChERS method and liquid chromatography tandem mass spectrometry (LC-MS/MS). The developed method was validated with average recoveries ranging from 63.10 to 102.10%, and relative standard deviations lower than or equal to 6.39% were obtained. Good linearity over the concentration range of 10-600 µg/kg dry matter (DM) was observed for the target alkaloids. The determination coefficients R2 calculated for each of the matrix calibration curves were greater than 0.99. The limits of detection and quantification were 5 µg/kg DM and 10 µg/kg DM, respectively. The reproducibility of the method was verified in three laboratories: all of the mean recoveries of 15 alkaloids were higher than 60%, and the relative standard deviations in alkaloids between laboratories were all less than 14.24%. The proposed method was applied to analyse 134 forage grass samples from the meadow steppe of Inner Mongolia to monitor toxic alkaloids. A significant difference in the frequency of contamination was found between different herbage species and different regions.

[Characteristics of light energy utilization of intercropping alfalfa/gramineae forage based on yield effect]. / 基于产量效应的间作紫花苜蓿/ç¦¾æœ¬ç§‘ç‰§è‰å ‰èƒ½åˆ©ç”¨ç‰¹å¾.

Based on field experiments in 2017-2019, we examined the characteristics, yield effect and regulatory mechanism of light energy utilization in alfalfa/gramineous forage grass intercropping. With monoculture of alfalfa, forage triticale (C3 plant), and forage maize (C4 plant) as control, we measured the yield effect, the effect of light energy utilization factor on yield formation, the characteristic difference and mechanism of light energy utilization under alfalfa/triticale and alfalfa/maize intercropping patterns. Results showed that land equivalent ratios of both intercropping patterns were all greater than 1, indicating that land utilization ratio and yield benefit of the two intercropping patterns were higher than that of monoculture, among which alfalfa/triticale intercropping pattern was the most promising one. The contribution of light energy utilization factors to yield was following an order of leaf area index (1.531) > net photosynthetic rate (0.882) > intercellular CO2 concentration (0.282) > transpiration rate (-0.229) > canopy opening (-0.291) > PAR interception rate (-0.681) > stomatal conductance (-0.751). Among them, leaf area index was not only one of the important indices to characterize photosynthetic capacity, but also an important component factor of forage crop yield aiming at harvesting nutrients. Therefore, among all factors of photosynthetic characteristics, net photosynthetic rate was the main factor affecting yield. The net photosynthetic rate of alfalfa, triticale and maize under intercropping showed the same pattern, and being different from that of monoculture. The main ways for intercropping to increase net photosynthetic rate included: triticale and maize increased net photosynthetic rate and yield by enhancing the carboxylation fixation capacity of CO2 and the utilization capacity of strong light, while alfalfa could improve its net photosynthetic rate and promote growth under low light, by increasing the content of chlorophyll b in functional leaves, changing chlorophyll composition and enhancing the collection and transmission of light energy.

Isolation of Intact Chloroplast for Sequencing Plastid Genomes of Five Festuca Species.

Isolation of good quality chloroplast DNA (cpDNA) is a challenge in different plant species, although several methods for isolation are known. Attempts were undertaken to isolate cpDNA from Festuca grass species by using available standard protocols; however, they failed due to difficulties separating intact chloroplasts from the polysaccharides, oleoresin, and contaminated nuclear DNA that are present in the crude homogenate. In this study, we present a quick and inexpensive protocol for isolating intact chloroplasts from seven grass varieties/accessions of five Festuca species using a single layer of 30% Percoll solution. This protocol was successful in isolating high quality cpDNA with the least amount of contamination of other DNA. We performed Illumina MiSeq paired-end sequencing (2 × 300 bp) using 200 ng of cpDNA of each variety/accession. Chloroplast genome mapping showed that 0.28%-11.37% were chloroplast reads, which covered 94%-96% of the reference plastid genomes of the closely related grass species. This improved method delivered high quality cpDNA from seven grass varieties/accessions of five Festuca species and could be useful for other grass species with similar genome complexity.

The Root Endophytic Fungi Community Structure of Pennisetum sinese from Four Representative Provinces in China.

Pennisetum sinese is a good forage grass with high biomass production and crude proteins. However, little is known about the endophytic fungi diversity of P. sinese, which might play an important role in the plant's growth and biomass production. Here, we used high throughput sequencing of the Internal Transcribed Spacer (ITS) sequences based on primers ITS5-1737 and ITS2-2043R to investigate the endophytic fungi diversity of P. sinese roots at the maturity stage, as collected from four provinces (Shaanxi province, SX; Fujian province, FJ; the Xinjiang Uyghur autonomous prefecture, XJ and Inner Mongolia, including sand (NS) and saline-alkali land (NY), China). The ITS sequences were processed using QIIME and R software. A total of 374,875 effective tags were obtained, and 708 operational taxonomic units (OTUs) were yielded with 97% identity in the five samples. Ascomycota and Basidiomycota were the two dominant phyla in the five samples, and the genera Khuskia and Heydenia were the most abundant in the FJ and XJ samples, respectively, while the most abundant tags in the other three samples could not be annotated at the genus level. In addition, our study revealed that the FJ sample possessed the highest OTU numbers (242) and the NS sample had the lowest (86). Moreover, only 22 OTUs were present in all samples simultaneously. The beta diversity analysis suggested a division of two endophytic fungi groups: the FJ sample from the south of China and the other four samples from north or northwest China. Correlation analysis between the environmental factors and endophytic fungi at the class level revealed that Sordariomycetes and Pucciniomycetes had extremely significant positive correlations with the total carbon, annual average precipitation, and annual average temperature, while Leotiomycetes showed an extremely significant negative correlation with quick acting potassium. The results revealed significant differences in the root endophytic fungi diversity of P. sinese in different provinces and might be useful for growth promotion and biomass production in the future.