Characterization and Pathogenicity of Pseudopestalotiopsis vietnamensis Causing Gray Blight of Wuyi Rock Tea(Camellia sinensis)in China and Specific Mechanisms of Disease Infection

Gray blight disease(GBD)causes significant losses in tea production in China.Although genes and biological processes involved in resistance to fungal disease in tea plants have been identified,specific mechanisms of the GBD infection process remain unknown.In this study,morphological and multi-gene(TEF-TUB-ITS)phylogenetic characteristics were used to identify isolate CLBB1 of Pseudopestalotiopsis vietnamensis.Pathogenicity tests confirmed that isolate CLBB1 from tea leaves caused GBD in the susceptible tea cultivar Wuyi Rock(Camellia sinensis var.sinensis cv.Shuixian).Spores began to germinate 24 h after infection(hai),and after 48 h,elongated fungal hyphae formed from a single conidium.Transcriptome analysis revealed that 482,517,and 369 genes were differentially expressed at 24,48,and 72 hai,respectively,in Wuyi Rock tea leaves.Isolate CLBB1 infection elicited phenotype-related responses and activated defense-related pathways,including plant–pathogen interaction,MAPK signaling,and plant hormone signal transduction,suggesting a possible mechanism underlying phenotype-based susceptibility to CLBB1.Thus,a new Ps.vietnamensis strain causing GBD in the tea cultivar‘Shuixian’was discovered in this study.Transcriptome analysis indicated that pathogen invasion activated chitin-related MAPK pathways and that tea plants required a hormone to restrict CLBB1.

Characteristic volatile components and transcriptional regulation of seven major tea cultivars(Camellia sinensis)in China

The volatiles in the young shoots of tea cultivars are the important material basis for the formation of tea aroma,but the cultivar-specific aroma and its molecular regulation are still lacking in research.In this study,the characteristic volatiles of seven tea cultivars in China were detected,and the results showed that the green tea cultivars'Fuding Dabaicha'(FDDB),'Longjing43'(LJ43),'Shuchazao'(SCZ),and'Baihaozao'(BHZ)were rich in(E)-3-hexenol,phenylethyl alcohol,phenylacetaldehyde,andβ-ionone.For oolong tea cultivars,the characteristic volatiles of'Tieguanyin'(TGY)were heptanal and eugenol,while the contents of(E)-β-ocimene,geraniol,and methyl salicylate were significantly increased in'Jinxuan'(JX).In addition,'Fujian Shuixian'(FJSX)has the highest content of esters,mainly jasmonolactone and dihydrojasmonolactone.Transcriptomic analysis showed that the different tea cultivars were significantly enriched in different levels of gene transcription in the three pathways related to aroma biosynthesis.Potential regulatory modules and genes of several characteristic volatiles were identified by WGCNA,among which CsbHLH(CsTGY12G0001520)may regulate the expression of CsTPS(CsTGY05G0001285)to directly affect the accumulation ofβ-caryophyllene in young shoots,while CsMYB(CsTGY01G0001203,CsTGY04G0001918,CsTGY06G0002545)may affect the synthesis of(Z)-3-hexenol and(E)-3-hexen-1-ol acetate by regulating the CsADH(CsTGY09G0001879).In addition,the transcription factors bHLH,WRKY,ERF,and MYB may be involved in the biosynthesis of linalool by regulating the expression of CsLIS/NES(CsTGY08G0001704,CsTGY08G0000359)genes individually or through interaction.These results revealed the characteristic volatiles and their key regulatory genes of seven tea cultivars,which will provide a theoretical basis for breeding and suitability research of tea cultivars.

Integrated transcriptome and metabolome provide insights into flavonoid biosynthesis in'P113',a new purple tea of Camellia tachangensis

Plants typically exhibit the purple phenomenon as a result of an increase in flavonoids and anthocyanins.A new tea germplasm'P113'was recently selected from Camellia tachangensis,which is purple in tender shoots.In the present study,integrated transcriptome and metabolome were used to analyze the flavonoid metabolite components and the genes involved in flavonoid biosynthesis in'P113'.A total of 86 flavonoid metabolites were identified,including 70 significantly differential metabolites(p<0.05)and they were enriched to the three metabolic pathways of ko00941,ko00942 and ko00944 through KEGG pathway analysis.A total of 136 flavonoid involved genes were obtained from transcriptomic study,of which 53 were significantly differentially expressed in developmental shoots.The correlation between metabolite profiling and transcriptome,transcriptome and protein interactions suggested that transcription factor MYB12 and glycosyltransferase UGT78D2 had a good facilitation on purple tender shoots.The metabolic pathways and potential genes that underlie the coloration of the shoots in'P113'are clarified in this study.It also lays the groundwork for identifying potential genes involved in color variation and offers a theoretical framework for the creation and use of distinctive genetic resources and the breeding of new cultivars.

Genome-Wide Identification of ABCC Gene Subfamily Members and Functional Analysis of CsABCC11 in Camellia sinensis

The ATP-binding cassette(ABC)transporter is a gene superfamily in plants.ATP-binding cassette subfamily C(ABCC)protein is a multidrug resistance-associated(MRP)transporter.They play various roles in plant growth,development,and secondary metabolite transport.However,there are few studies on ABCC transporters in tea plants.In this study,genome-wide association study(GWAS)analysis of epigallocatechin gallate(EGCG)content in 108 strains of Kingbird revealed that CsABCCs may be involved in EGCG transport.We identified 25 CsABCC genes at the genomic level of the tea plant,their phylogenetic tree,gene structure,targeted miRNA and other bioinformatics were analyzed.The expression patterns of CsABCCs in eight different tissues and abiotic stress indicate that they have potential roles in regulating the growth,development,and defense of tea plants.The correlation analysis revealed that the expression of the CsABCC11 gene was closely related to the EGCG content in tea buds of 108 strains of the Kingbird,and the subcellular localization experiments in tobacco showed that CsABCC11 protein was localized on the plasma membrane.The virus-induced gene silencing(VIGS)strategy in tea plants further verified that CsABCC11 was involved in EGCG accumulation.Our study laid a foundation for studying the biological function of CsABCC and provided a new candidate molecular marker gene for further EGCG-related variety breeding,which will be of great interest to breeders.

Evaluation of the Resistance of Different Tea Cultivars to Tea Aphids by EPG Technique

In order to investigate the resistance of tea plant to tea aphid, the feeding behavior of tea aphids on six different tea cultivars was monitored by （EPG） technique. The result showed that the duration of El, E2 as well as （E I＋E2） which was feeding waveforrns of Toxoptera aurantii （Boyer） in tea phloem was significantly different by variance analysis （P〈0.05） and so did the duration of the probing of T. aurantii in all the tea tissues. And the six tea cultivars were classified into three groups with cluster analysis, based on the six major parameters which were durations of np, C, El, E2, F, and G waveforms, then the six major parameters of these three groups were detected by variance analysis, and the rank of six different tea cultivars＇ resistance to T. aurantii from strong to weak was Zhongcha 108, Sucha 1, Anjibaicha, Longjing 43, Xicha 5, and Sucha 120. As this result was in accord with the investigation into the tea field, we concluded that EPG technique was one of the most important means to examine tea plant resistance to T. aurantii and the durations of waveform El, E2 and （El＋E2） were important parameters for evaluating resistance of tea plants to tea aphids.

Genome-level diversification of eight ancient tea populations in the Guizhou and Yunnan regions identifies candidate genes for core agronomic traits

The ancient tea plant,as a precious natural resource and source of tea plant genetic diversity,is of great value for studying the evolutionary mechanism,diversification,and domestication of plants.The overall genetic diversity among ancient tea plants and the genetic changes that occurred during natural selection remain poorly understood.Here,we report the genome resequencing of eight different groups consisting of 120 ancient tea plants:six groups from Guizhou Province and two groups from Yunnan Province.Based on the 8,082,370 identified high-quality SNPs,we constructed phylogenetic relationships,assessed population structure,and performed genome-wide association studies(GWAS).Our phylogenetic analysis showed that the 120 ancient tea plants were mainly clustered into three groups and five single branches,which is consistent with the results of principal component analysis(PCA).Ancient tea plants were further divided into seven subpopulations based on genetic structure analysis.Moreover,it was found that the variation in ancient tea plants was not reduced by pressure from the external natural environment or artificial breeding(nonsynonymous/synonymous=1.05).By integrating GWAS,selection signals,and gene function prediction,four candidate genes were significantly associated with three leaf traits,and two candidate genes were significantly associated with plant type.These candidate genes can be used for further functional characterization and genetic improvement of tea plants.

Population Structure Analysis and Genome-Wide Association Study of Tea(Camellia sinensis(L.)Kuntze)Germplasm in Qiannan,China,Based on SLAF-Seq Technology

Duyun Maojian tea is a famous tea in China.In this study,the specific-locus amplified fragment(SLAF)sequencing method was used to analyze the population structure and conduct a genome-wide association study(GWAS)of 2 leaf traits of 123 tea plants in Qiannan,China.A total of 462,019 SLAF tags and 11,362,041 single-nucleotide polymorphism(SNP)loci were obtained.The results of phylogenetic tree analysis,cluster analysis,and principal component analysis showed that 123 tea germplasms were clustered into three groups,and the heterozygosity rates for Groups I,II,and II were 0.206,0.224,and 0.34,respectively.Generally,tea germplasms in a production area are clustered in a group,indicating that tea germplasms in different production areas have certain genetic diversity.The traditional Duyun Maojian tea core production areas,TS and DC-SJ,are clustered into Group I and Group II respectively,while the ZY production area is relatively independent in Group III.Furthermore,based on GWAS analysis,11 candidate genes related to leaf apex and 7 candidate genes related to leaf shape were obtained.This study clarified the genetic relationship among eight Duyun Maojian tea production areas and obtained candidate genes related to leaf apex and leaf shape development.The results showed that population structure and candidate genes are an effective basis for the breeding of Duyun Maojian tea germplasm.

Identification of miRNAs and target genes regulating catechin biosynthesis in tea (Camellia sinensis)

Micro RNAs（miRNAs） are endogenous non-protein-coding small RNAs that play crucial and versatile regulatory roles in plants. Using a computational identification method, we identified 55 conserved miRNAs in tea（Camellia sinensis） by aligning miRNA sequences of different plant species with the transcriptome library of tea strain 1005. We then used quantitative real-time PCR（qRT-PCR） to analyze the expression of 31 identified miRNAs in tea leaves of different ages, thereby verifying the existence of these miRNAs and confirming the reliability of the computational identification method. We predicted which miRNAs were involved in catechin synthesis using ps RNAtarget Software based on conserved miRNAs and catechin synthesis pathway-related genes. Then, we used RNA ligase-mediated rapid amplification of c DNA ends（RLM-RACE） to obtain seven miRNAs cleaving eight catechin synthesis pathway-related genes including chalcone synthase（CHS）, chalcone isomerase（CHI）, dihydroflavonol 4-reductase（DFR）, anthocyanidin reductase（ANR）, leucoanthocyanidin reductase（LAR）, and flavanone 3-hydroxylase（F3H）. An expression analysis of miRNAs and target genes revealed that miR529d and miR156 g-3 p were negatively correlated with their targets CHI and F3H, respectively. The expression of other miRNAs was not significantly related to their target genes in the catechin synthesis pathway. The RLM-RACE results suggest that catechin synthesis is regulated by miRNAs that can cleave genes involved in catechin synthesis.

Controlled-Release of Plant Volatiles:New Composite Materials of Porous Carbon-Citral and Their Fungicidal Activity against Exobasidium vexans

Citral(Eo)exhibits excellent fungicidal activities.However,it is difficult to maintain long-term fungicidal activity due to its strong volatility.Herein,a controlled-release strategy by using biomass-derived porous carbon(BC)was developed to overcome the drawback of Eo.New composite materials were prepared by loading Eo on tea stem porous carbon(BC@Eo),and their controlled-release fungicidal activity against Exobasidium vexans was assessed.BC with a large specific surface area of 1001.6 m2/g and mesoporous structure was fabricated through carbonization tempera-ture of 700℃.The BC@Eo materials were characterized using Fourier-transform infrared spectroscopy and X-ray powder diffraction.The results suggested that chemical and physical interactions occurred in BC@Eo.The Eo release profile suggested a biphasic pattern with an initial fast release on days 1–14 and a subsequent controlled phase on days 14–30.The in vitro cumulative release percentage of Eo from BC@Eo was 51%during one month,and this result was significantly lower than that from free Eo(cumulative release percentage of Eo of 82%in one week).The anti-fungal activities of Eo and BC@Eo against E.vexans were determined using the inhibition zone method.The results indicated that Eo and BC@Eo formed large inhibition zones of 19.66±0.79 and 21.92±0.77 mm,respectively.The influence on the hyphal structure of E.vexans was observed by scanning electron microscopy on day 30.The hyphal structure of E.vexans treated with BC@Eo was more shrunken than that treated with Eo at 30 days,suggesting that BC@Eo prolongs the fungicidal activity against E.vexans.This study demonstrated that the encapsulation of Eo in BC for developing the BC@Eo materials could be a promising strategy to inhibit volatility and maintain the fungicidal activity of Eo and provide a potential alternative for the reuse of abundant tea biomass waste resources.

Genome-Wide Characterization of the Cellulose Synthase Gene Superfamily in Tea Plants(Camellia sinensis)

The cellulose synthase gene superfamily,including Cellulose synthase A(CesA)and cellulose synthase-like(Csl)gene families,is responsible for the synthesis of cellulose and hemicellulose,respectively.The CesA/Csl genes are vital for abiotic stress resistance and shoot tenderness regulation of tea plants(Camellia sinensis).However,the CesA/Csl gene family has not been extensively studied in tea plants.Here,we identified 53 CsCesA/Csl genes in tea plants.These genes were grouped into five subfamilies(CsCesA,CsCslB,CsCslD,CsCslE,CsCslG)based on the phylogenetic relationships with Arabidopsis and rice.The analysis of chromosome distribution,gene structure,protein domain and motif revealed that most genes in CsCesA,CsCslD and CsCslE subfamilies were conserved,whereas CsCslB and CsCslG subfamily members are highly diverged.The transcriptome analysis showed that most CsCesA/Csl genes displayed tissue-specific expression pattern.In addition,members of CsCslB4,CsCesA1/3/6,CsCslB3/4,CsCslD3,CsCslE1 and CsCslG2/3 subfamilies were up-regulated under drought and cold stresses,indicating their potential roles in regulating stress tolerance in tea plants.Furthermore,the expression levels of CsCslG2_6 and CsCslD3_5 in different tissues and cultivars,respectively,were positively correlated with the cellulose content that is negatively related with shoot tenderness.Thus,these two genes were speculated to be involved in the regulation of shoot tenderness in tea plants.Our findings may help elucidate the evolutionary relationships and expression patterns of the CsCesA/Csl genes in tea plants,and provide more candidate genes responsible for stress tolerance and tenderness regulation in tea plants for future functional research.

Canopy Cultivation Technology and Benefit Analysis of Tea Tree Canopy in Preserved Tea Plantations

The present study introduced the concept and characteristics of preserved tea plantation,and analyzed application and benefit of stereoscopic trimming and pruning technology in the cultivation of tea tree canopy in preserved tea plantations.The results show that the stereoscopic trimming and pruning technology of tea trees has more advantages in production efficiency than other trimming and pruning techniques in terms of yield and output value,and can better exert the regeneration ability of tea branches.The grade of branching is increased from grade 3-4 before unpruning to grade 9-13(3-4 times higher),the length of internode is increased from 1-2 cm to 3-7 cm(increased 3-4 times),the picking area is increased from 0.9 m×1.1 m to 2.8 m×4.2 m(an increase of nearly 10 times),accordingly achieving the best production and economic benefits required in tea production.

Cloning and Function Identification of a Phytoene Desaturase Gene from Eucommia ulmoides

The phytoene desaturase(PDS)encodes a crucial enzyme in the carotenoid biosynthesis pathway.Silencing or inhibiting PDS expression leads to the appearance of mottled,chlorosis,or albino leaves.In this study,the CDS sequence of EuPDS(Eucommia ulmoides Phytoene Desaturase)was first cloned and then PDS was silenced in Nicotiana benthamiana.Result showed the expression level of EuPDS in leaves was higher than that in the roots and stems.In N.benthamiana leaves,which were treated by Agrobacterium for 24 h,photo-bleaching was shown on the fresh leaves one week after injection and the transcript level of PDS was down-regulated during the period of emersion.This suggested that EuPDS could silence PDS of N.benthamiana,so as to cause the phenotype of leaf whitening.PDS is the main reporter gene involved in virus-induced gene silencing(VIGS).This study offered molecular evidence for identifying PDS gene involved in Carotenoid’s biosynthesis pathway and the regulation networks in E.ulmides.It also laid a useful foundation for study on leaf discoloration mechanism of other woody plants.

Overexpression of a sugarcane ScCaM gene negatively regulates salinity and drought stress responses in transgenic Arabidopsis thaliana

Calmodulin(CaM)proteins play a key role in signal transduction under various stresses.In the present study,the effects of a sugarcane ScCaM gene(NCBI accession number:GQ246454)on drought and salt stress tolerance in transgenic Arabidopsis thaliana and Escherichia coli cells were evaluated.The results demonstrated a significant negative role of ScCaM in the drought and salt stress tolerance of transgenic lines of A.thaliana,as indicated by the phenotypes.In addition,the expression of AtP5CS and AtRD29A,two genes tightly related to stress resistance,was significantly lower in the overexpression lines than in the wild type.The growth of E.coli BL21 cells expressing ScCaM showed weaker tolerance under mannitol and NaCl stress.Taken together,this study revealed that the ScCaM gene plays a negative regulatory role in both mannitol and NaCl stresses,and it possibly exerts protective mechanisms common in both prokaryotes and eukaryotes under stress conditions.

Overexpression ofβ-1,4-Glucanase Gene EuEG1 Improves Micrografting of Eucommia ulmoides

Adventitious root formation poses a major constraint on the tissue culture and genetic transformation of Eucommia ulmoides.Micrografting offers a new method for the transplantation of genetic transformation,and its success depends on the formation of graft unions.This study used transgenic rootless test-tube seedlings as scions and seedlings from seed as rootstocks during micrografting to avoid the rooting issues that occur during tissue culture and to investigate the role of the EuEG1 gene in the graft healing process.We found that the EuEG1 gene is a vital regulator of graft,and its overexpression contributes to the survival of Eucommia ulmoides micrografting.The EuEG1 gene transgenic plants(TP)used as scions for micrografting presented a significantly higher survival rate than the wild type(WT)and empty vector(EV)regenerated scions.During the grafting healing process,the expression of the EuEG1 gene was higher during the period of callus proliferation,suggesting that the EuEG1 gene was involved in the graft healing process.Histological observation revealed that more calluses tissue appeared at the junction of transgenic scions,and the connection with the rootstock was stronger,which benefits wound healing.These results provide new insights into Eucommia ulmoides micrografting and indicate that the EuEG1 gene can promote wound healing and improve the micrografting survival rate.

An NBS-LRR-encoding gene CsRPM1 confers resistance to the fungus Colletotrichum camelliae in tea plant

Nucleotide binding site,leucine-rich repeat(NBS-LRR)proteins are the main types of disease resistance proteins in plants,which can perceive plant pathogens.Anthracnose,caused by the fungus Colletotrichum camelliae,is one of the most severe diseases in tea plant.Here,we identified an NBS-LRR-encoding gene,CsRPM1,probably conferring resistance of tea plant to C.camelliae.Phylogenetic analysis showed that CsRPM1 was clustered with RPM1 in Arabidopsis and grouped into CC-NBS-LRR(CNL).It contained a signal peptide,a NB-ARC domain,and multiple LRR motifs.RNA-seq and qRT-PCR analysis showed that the transcript level of CsRPM1 was significantly up-regulated after inoculation with C.camelliae.Transiently silencing of CsRPM1 in tea leaves comprised the resistance to C.camelliae,indicating that CsRPM1 was required for plant defense against the pathogen.The subcellular localization showed that CsRPM1 protein was localized in the nucleus,cytoplasm,and cell membrane.Furthermore,the promoter region of CsRPM1 gene contained MeJA-responsive elements,and the expression of CsRPM1 was induced by exogenous methyl jasmonate,suggesting that CsRPM1 gene may be closely related to JA signaling pathway.A total of 17 transcription factors might be responsible for the expression of CsRPM1.Our data indicates that CsRPM1 is required for disease resistance to C.camelliae in tea plant.The characterization of this disease resistance gene sheds light on NLR protein function in tea plant and may facilitate breeding to control the severe anthracnose.

Characterization of eating quality and starch properties of two Wx alleles japonica rice cultivars under different nitrogen treatments

To understand the effect of nitrogen(N) fertilizer on rice(Oryza sativa L.) eating and cooking quality(ECQ). Here, we investigated the ECQ attributes, physicochemical foundation of ECQ, and amylopectin fine structure of two Waxy(Wx) alleles japonica rice cultivars Nanjing 9108(NJ9108) and Huaidao 5(HD5) under four N rates(0, 150, 300, and 450 kg ha^-1). Sensory and pasting properties of the two cultivars varied depending on N rates. Compared with the control(0 kg ha^-1), the overall eating quality and sensory value were significantly decreased under the N rates of 300 and 450 kg ha^-1. Further, conventional descriptive analysis showed that the stickiness and retrogradation of cooked rice were significantly decreased. These results indicated that application of N fertilizer seems to affect the texture of cooked rice, causing it to be less sticky, lowering its retrogradation, and consequently reducing its palatability. Results from rapid visco analyzer(RVA) revealed that the peak and breakdown viscosities significantly decreased, while the setback viscosity and peak time increased under the N rates of 300 and 450 kg ha^-1. However, no significant difference was observed when the N rate was 150 kg ha^-1, indicating that less N fertilization can maintain rice ECQ. As the N rate increasing, protein content increased, whereas apparent amylose content, starch content, and gel consistency almost unchanged. Interestingly, compared with the control, under N treatments, the percentage of short amylopectin branches in NJ9108 was decreased, but increased in HD5, as controlled by amylopectin synthesis-related genes. Notably, SSI and BEIIb were down-regulated in NJ9108, whereas BEIIb was up-regulated in HD5. Thus, the palatability of both rice cultivars was significantly decreased under excessive N fertilization as a consequence of reduced stickiness and retrogradation of the cooked rice, which might have resulted from an elevated protein content and altered amylopectin fine structure. In addition, amylopectin synthesis appeared to be affected by N fertilizer and the genotype of the rice cultivar.

Isolation and Characterization of Fucoidans from Five Brown Algae and Evaluation of Their Antioxidant Activity

In this study, we evaluated the chemical property and antioxidant activity of fucoidans isolated from brown algae, Laminaria japonica(LJF), Lessonia nigrescens(LNF), Lessonia trabeculata(LTF), Ascophyllum mackaii(AMF), and Ecklonia maxima(EMF). LJF was less in sulfate content(14.16%) and more in galactose and mannose content(1.08 and 0.68) than the documented early. EMF contained 20%–30% of sulfate and fucose, 0.97 in molar ratio which was lower than that of sulfate to other four fucoidans(1.21–1.41). AMF(162 kDa) and EMF(150 kDa) were the first two largest in molecular weight, which were followed by LJP(126 kDa), LNF(113 kDa) and LTF(105 kDa). The fucoidans isolated these algae showed a wide range of antioxidant activity in vitro. It was found that the reducing power of the isolated fucoidans was positively correlated with their sulfate content and molecular weight. In addition, LNF and LTF at low concentrations exhibited high superoxide and hydroxyl radical scavenging activity. This demonstrated that low molecular weight fucoidans may perform a high antioxidant activity.

Storage Stability in Reversion Mutation of a Rice Line Devoid of LOX-1, 2 Acquired by Ion Beam Irradiation

The effect of absence of lipoxygenase isoenzyme （LOX） on storage stability was investigated. Rice mutant 1297 without lipoxygenase isoenzyme-1 （LOX-1） or lipoxygenase isoenzyme-2 （LOX-2） generated by ion beam irradiation from Wanjian2090 and reversion mutant RM1297 with LOX-1 and LOX-2 were subjected to an accelerated-aging experiment. Shanyou63 （with LOX-1 and LOX-2 ） served as control. Results showed that the germination and dehydrogenase activity decreased while the electrical conductivity and free fatty acid content increased in all varieties with accelerated aging. In 1297 that lacked LOX-1 and 2, there were slight changes in germination, dehydrogenase activity, membrane permeability and free fatty acid content during the thirty-day accelerated-aging experiment. But in varieties with LOX-1 and LOX-2, significant changes were observed, suggesting that LOX-1, 2 might be a definite factor which influenced seed lifespan. This study also indicates that ion beam irradiation may be used as mutagen to generate mutant and reversion mutants for biological study and could become a new direction in ion beam application.

Nutrient value of wild fodder species and the implications for improving the diet of mithun (Bos frontalis) in Dulongjiang area, Yunnan Province, China

Wild fodder plants are valuable because they provide an important livestock feed resource globally,especially for smallholder farmers,and have important roles in natural resource management.In-depth knowledge of wild forage plants can motivate local people for feed resource and habitat conservation of threatened herbivores such as Mithun(Bos frontalis).Mithun occur in small patches in the mountains of Dulongjiang,nearby villagers domesticated this animal but left animal to freely graze in the mountains.Many fodder plants occur in these mountains,however,little is known about their nutritional value.We conducted an ethnobotanical survey to document important wild fodder plants consumed by mithun in the Dulongjiang Township.The nutritional content of 21 highly mentioned wild fodder plants in an ethnobotanical survey was examined.Laboratory analysis showed that Fagopyrum dibotrys were the fodder species with the highest crude protein(CP)content(26.89%),followed by Polygonum molle(21.88%)and Hydrangea longipes(21.12%).Synthesis of relative feed value index and grey relational grade,P.molle,H.longipes and Tetrastigma obtectum were ranked the top three nutritional fodders.There was a significant difference between 21 species on their in vitro digestibility and the most highly digestible fodder species was Elatostema hookerianum.Linear model analysis on relationship between frequency of citation of 21 wild forage plants by local farmers and their nutrient composition showed that the frequency was significantly positively correlated with the nutritional value of the feed(R2?0.28,P<0.05).We concluded that these species have high nutritional values to improve mithun production in integrated crop-livestock systems.Fodder species or mixtures of species with useful nutritional characters could be cultivated to improve livestock productivity,habitat conservation including that of mithun and wild forage resource management.

Cloning and Characterization of EuGID1 in Eucommia ulmoides Oliver

Gibberellic acid controlled the key developmental processes of the life cycle of landing plants,and regulated the growth and development of plants.In this study,a novel gibberellin receptor gene EuGID1 was obtained from Eucommia ulmoides Oliver.The cDNA of EuGID1 was 1556 bp,and the open reading frame was 1029 bp,which encoded 343 amino acids.EuGID1 had the homology sequence with the hormone-sensitive lipase family.Amino acid sequence alignment confirmed EuGID1 protein had the highest homology with the GID1 protein of Manihot esculenta.EuGID1 was located in the nucleus and cell membrane and had expression in four plant organs.Overexpression of EuGID1 in transgenic Arabidopsis plants promoted plant elongation and increased siliques yield.