摘要
By investigating the contamination status and predicting the exposure risk of mycotoxin in Coicis Semen, we aim to provide guidance for the safety supervision of Chinese medicinal materials and the formulation(revision) of mycotoxin limit standards. The content of 14 mycotoxins in the 100 Coicis Semen samples collected from five major markets of Chinese medicinal materials in China was determined by UPLC-MS/MS. The probability evaluation model based on Monte Carlo simulation method was established after Chi-square test and One-way ANOVA of the sample contamination data. Health risk assessment was performed on the basis of margin of exposure(MOE) and margin of safety(MOS). The results showed that zearalenone(ZEN), aflatoxin B_1(AFB_1), deoxynivalenol(DON), sterigmatocystin(ST), and aflatoxin B_2(AFB_2) in the Coicis Semen samples had the detection rates of 84%, 75%, 36%, 19%, and 18%, and the mean contamination levels of 117.42, 4.78, 61.16, 6.61, and 2.13 μg·kg~(-1), respectively. According to the limit standards in the Chinese Pharmacopoeia(2020 edition), AFB_1, AFs and ZEN exceeded the standards to certain extents, with the over-standard rates of 12.0%, 9.0%, and 6.0%, respectively. The exposure risks of Coicis Semen to AFB_1, AFB2, ST, DON, and ZEN were low, while 86% of the samples were contaminated with two or more toxins, which needs more attention. It is suggested that the research on the combined toxicity of different mycotoxins should be strengthened to accelerate the cumulative exposure assessment of mixed contaminations and the formulation(revision) of toxin limit standards.
Subject(s)
Humans , Mycotoxins/analysis , Coix , Aflatoxin B1/analysis , Chromatography, Liquid/methods , Food Contamination/analysis , Tandem Mass Spectrometry/methods摘要
This paper aimed to study the role of asparagine endopeptidase(AEP) gene in the biosynthesis mechanism of cyclic peptide compounds in Pseudostellaria heterophylla. The transcriptome database of P. heterophylla was systematically mined and screened, and an AEP gene, tentatively named PhAEP, was successfully cloned. The heterologous function verification by Nicotiana benthamiana showed that the expression of the gene played a role in the biosynthesis of heterophyllin A in P. heterophylla. Bioinformatics analysis showed that the cDNA of PhAEP was 1 488 bp in length, encoding 495 amino acids with a molecular weight of 54.72 kDa. The phylogenetic tree showed that the amino acid sequence encoded by PhAEP was highly similar to that of Butelase-1 in Clitoria ternatea, reaching 80%. The sequence homology and cyclase active site analysis revealed that the PhAEP enzyme may specifically hydrolyse the C-terminal Asn/Asp(Asx) site of the core peptide in the HA linear precursor peptide of P. heterophylla, thereby participating in the ring formation of the linear precursor peptide. The results of real-time quantitative polymerase chain reaction(RT-qPCR) showed that the expression level of PhAEP was the highest in fruits, followed by in roots, and the lowest in leaves. The heterophyllin A of P. heterophylla was detected in N. benthamiana that co-expressed PrePhHA and PhAEP genes instantaneously. In this study, the PhAEP gene, a key enzyme in the biosynthesis of heterophyllin A in P. heterophylla, has been successfully cloned, which lays a foundation for further analysis of the molecular mechanism of PhAEP enzyme in the biosynthesis of heterophyllin A in P. heterophylla and has important significance for the study of synthetic biology of cyclic peptide compounds in P. heterophylla.
Subject(s)
Genes, vif , Phylogeny , Plant Leaves/genetics , Peptides, Cyclic , Cloning, Molecular , Caryophyllaceae/genetics摘要
This study aimed to establish a method for synchronous detection of 14 mycotoxins in Pseudostellariae Radix and investigate its contamination with mycotoxins, so as to provide technical guidance for monitoring the quality of Chinese medicinal materials and medication safety. The sample was extracted with 80% acetonitrile in an oscillator for 1 h, purified using the modified QuEChERS purifying agent(0.1 g PSA + 0.3 g C_(18) + 0.3 g MgSO_4), and separated on a Waters HSS T3 chromatographic column(2.1 mm×100 mm, 1.8 μm). The gradient elution was carried out with 0.1% formic acid in water and acetonitrile, followed by the scanning in the multi-reaction monitoring(MRM) mode and the analysis of mycotoxin contamination in 26 Pseudostellariae Radix samples. The recovery rates of the established method were within the range of 82.17%-113.6%, with the RSD values less than 7% and the limits of quantification(LOQ) being 0.019-0.976 μg·kg~(-1). The detection rate of 14 mycotoxins in 26 batches of medicinal materials was 53.85%. The detection rate of sterigmatocystin(ST) was the highest, followed by those of zearalenone(ZEN), aflatoxin G_2(AFG_2), fumonisin B_1(FB_1), HT-2 toxin, and nivalenol(NIV). Their respective detection rates were 38.46%, 26.92%, 23.08%, 11.54%, 11.54%, and 7.69%, with the pollution ranges being 1.48-69.65, 0.11-31.05, 0.11-0.66, 0.28-0.83, 20.86-42.56, and 0.46-1.84 μg·kg~(-1), respectively. The established method for the detection of 14 mycotoxins is accurate, fast and reliable. The research results have very important practical significance for guiding the monitoring and prevention and control of exogenous fungal contamination of Chinese medicinal materials.
Subject(s)
Aflatoxins/analysis , Chromatography, High Pressure Liquid/methods , Drug Contamination , Food Contamination/analysis , Mycotoxins/analysis , Plant Roots/chemistry , Tandem Mass Spectrometry/methods摘要
Due to the special biological characteristics, Gastrodia elata suffers from high resource consumption and low utilization rate in modern agricultural production, which significantly block the green and healthy development of this industry. Based on the theory and technology in ecological cultivation of Chinese medicinal materials, this study analyzed the challenges in ecological cultivation of G. elata, such as waste of fungus material, a few cultivation modes available, continuous cropping obstacles, frequent occurrence of diseases, and poor stability of ecological structure. According to the production practice, the following suggestions were proposed for ecological cultivation of G. elata: following the principle of environmental protection and no pollution, selecting suitable habitats to yield high-quality medicinal materials, committing to green control of diseases and pests, upgrading industrial structure to maximize the benefits, establishing a sound mechanism for protecting the genetic diversity of wild G. elata, carrying out simulative habitat cultivation to improve medicinal material quality, adopting science-based planning of fungus resources to relieve forestry pressure, enhancing the recycling and utilization of fungus materials, and applying diversified cultivation modes to improve the stability of ecological structure. The result is expected to provide a reference for the quality development of G. elata industry.
Subject(s)
Agriculture , Gastrodia/chemistry , Plants, Medicinal/chemistry摘要
Tuber rot has become a serious problem in the large-scale cultivation of Gastrodia elata. In this study, we compared the resistance of different ecotypes of G. elata to tuber rot by field experiments on the basis of the investigation of G. elata diseases. The histological observation and transcriptome analysis were conducted to reveal the resistance differences and the underlying mechanisms among different ecotypes. In the field, G. elata f. glauca had the highest incidence of tuber rot, followed by G. elata f. viridis, and G. elata f. elata and G. elata f. glauca×G. elata f. elata showed the lowest incidence. Tuber rot showcased obvious plant source specificity and mainly occurred in the buds and bottom of G. elata plants. After infection, the pathogen spread hyphae in host cortex cells, which can change the endophytic fungal community structure in the cortex and parenchyma of G. elata. G. elata f. glauca had thinner lytic layer and more sugar lumps in the parenchyma than G. elata f. elata. The transcription of genes involved in immune defense, enzyme synthesis, polysaccharide synthesis, carbohydrate transport and metabolism, hydroxylase activity, and aromatic compound synthesis had significant differences between G. elata f. glauca and G. elata f. elata. These findings suggested that the differences in resis-tance to tuber rot among different ecotypes of G. elata may be related to the varied gene expression patterns and secondary metabolites. This study provides basic data for the prevention and control of tuber rot and the improvement of planting technology for G. elata.
Subject(s)
Ecotype , Gastrodia/microbiology , Gene Expression Profiling , Plant Tubers/genetics摘要
Brown rot is a common disease in the cultivation and production of Gastrodia elata, but its pathogens have not been fully revealed. In this study, the pathogenic fungi were isolated and purified from tubers of 77 G. elata samples with brown rot. Pathogens were identified by the pathogenicity test and morphological and molecular identification. The pathogenicity of each pathogen and its inhibitory effects on Armillaria gallica were compared. The results showed that 119 strains of fungi were isolated from tubers of G. elata infected with brown rot. Among them, the frequency of separation of Ilyonectria fungi was as high as 42.01%. The pathogenicity test showed that the pathogenicity characteristics of six strains of fungi were consistent with the natural symptoms of brown rot in G. elata. The morphological and molecular identification results showed that the six strains belonged to I. cyclaminicola and I. robusta in the Nectriaceae family of Sordariomycetes class, respectively. Both types of fungi could produce pigments, conidia, and chlamycospore, and the growth rate of I. cyclaminicola was significantly higher than that of I. robusta. The comparison of pathogenicity showed that the spots formed by I. cyclaminicola inoculation were significantly larger than those of I. robusta inoculation, suggesting I. cyclaminicola was superior to I. robusta in pathogenicity. The results of confrontation culture showed that I. cyclaminicola and I. robusta could signi-ficantly inhibit the germination and cordage growth of A. gallica. A. gallica also inhibited the growth of pathogens, and I. cyclaminicola was less inhibited as compared with I. robusta. The results of this study revealed for the first time that I. cyclaminicola and I. robusta were the pathogens responsible for G. elata brown rot.
Subject(s)
Fungi , Gastrodia , Plant Tubers , Spores, Fungal , Virulence摘要
Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.
Subject(s)
Agaricales , Gastrodia/genetics , Protoplasts , Symbiosis/genetics , Transformation, Genetic摘要
Zearalenone(ZEN) is a mycotoxin produced by Fusarium, possessing estrogen-like effects, carcinogenicity, and multiple toxicities. To seek more efficient and practical agents for biological detoxification and broaden their application, this study isolated 194 bacterial strains from the moldy tuberous root of Pseudostellaria heterophylla, which were co-cultured with ZEN. An efficient ZEN-degrading strain H4-3-C1 was screened out by HPLC and identified as Acinetobacter calcoaceticus by morphological observation and molecular identification. The effects of culture medium, inoculation dose, culture time, pH, and temperature on the degradation of ZEN by H4-3-C1 strain were investigated. The mechanism of ZEN degradation and the degrading effect in Coicis Semen were discussed. The degradation rate of 5 μg·mL~(-1) ZEN by H4-3-C1 strain was 85.77% in the LB medium(pH 6) at 28 ℃/180 r·min~(-1) for 24 h with the inoculation dose of 1%. The degradation rate of ZEN in the supernatant of strain culture was higher than that in the intracellular fluid and thalli. The strain was inferred to secret extracellular enzymes to degrade ZEN. In addition, the H4-3-C1 strain could also degrade ZEN in Coicis Semen. If the initial content of ZEN in Coicis Semen was reduced from 90 μg·g~(-1) to 40.68 μg·g~(-1), the degradation rate could reach 54.80%. This study is expected to provide a new strain and application technology for the biological detoxification of ZEN in food processing products and Chinese medicinal materials.
Subject(s)
Bacteria , Fusarium , Mycotoxins , Temperature , Zearalenone摘要
Fusarium is the major pathogen of root rot of Pseudostellaria heterophylla. This study aims to explain the possible distribution of Fusarium species and the contamination of its toxin-chemotypes in tuberous root of P. heterophylla. A total of 89 strains of fungi were isolated from the tuberous root of P. heterophylla. Among them, 29 strains were identified as Fusarium by ITS2 sequence, accounting for 32.5%. They were identified as five species of F. avenaceum, F. tricinctum, F. fujikuroi, F. oxysporum, and F. graminearum based on β-Tubulin and EF-1α genes. LC-MS/MS detected 18, 1, and 5 strains able to produce ZEN, DON, and T2, which accounted for 62.1%, 3.4%, and 17.2%, respectively. Strain JK3-3 can produce ZEN, DON, and T2, while strains BH1-4-1, BH6-5, and BH16-2 can produce ZEN and T2. PCR detected six key synthase genes of Tri1, Tri7, Tri8, Tri13, PKS14, and PKS13 in strain JK3-3, which synthesized three toxins of ZEN, DON, and T2. Four key synthase genes of Tri8, Tri13, PKS14, and PKS13 were detected in strains BH1-4-1, BH6-5, and BH16-2, which were responsible for the synthesis of ZEN and T2. The results showed that the key genes of toxin biosynthesis were highly correlated with the toxins produced by Fusarium, and the biosynthesis of toxin was strictly controlled by the genetic information of the strain. This study provides a data basis for the targeted prevention and control of exo-genous mycotoxins in P. heterophylla and a possibility for the development of PCR for rapid detection of toxin contamination.
Subject(s)
Caryophyllaceae , Chromatography, Liquid , Fusarium/genetics , Mycotoxins , Tandem Mass Spectrometry摘要
Objective:To study the microbial community composition and diversity of brown-rot Gastrodia elata and its surface soil,in order to explain the relationship between brown-rot G. elata and soil microflora in G. elata planting process and provide theoretical basis for revealing the reasons of G. elata brown-rot disease. Method:Used internal transcribed spacer region(ITS) and 16S rDNA high-throughput sequencing technologies to detect the microbial diversity,community structure composition and community structure similarity of fungi and bacteria in healthy tuber,Brown-rot tuber,healthy soil and Brown-rot soil. Result:Compared with health groups,the number of fungi and bacteria operational taxonomic units(OTUs) was increased in brown-rot G. elata and its soil, and the abundance and diversity of fungi and bacteria in brown-rot G. elata soil were significantly decreased. The diversity of fungi in the tubers of brown-rot G. elata was significantly reduced,while the diversity of bacteria was significantly decreased. At the genus level, Mortierella was dominant fungi genus in healthy tuber and healthy soil,which was reduced 7.62% and 15.75% respectively in brown-rot tuber and brown-rot soil. And the dominant bacteria genus was Bradyrhizobium and Burkholderia-Paraburkholderia respectively. Ilyonectria was dominant fungi genus in brown-rot tuber and brown-rot soil,the dominant bacteria genus was Serratia and Bradyrhizobium respectively. Conclusion:The fungal flora in the tuber of brown-rot G. elata had a very high degree of similarity to that in the surrounding soil. These results indicated that the change of soil microbial fungal community caused the occurrence of G. elata brown-rot disease to a certain extent. And the pathogenic fungal Ilyonectria was dominant genus in fungi community of brown-rot tuber and brown-rot soil. Ilyonectria may have the main G. elata brown-rot disease pathogen.
摘要
Objective:To explore the effects of high temperature stress on the growth characteristics of different Armillaria strains,and to provide guidance for screening excellent Armillaria strains with high-temperature resistance. Method:14 strains of Armillaria from different G. elata producing areas were used as experimental materials to observe the growth characteristics and conduct phenotypic classification for the strains. rDNA-IGS sequence analysis was used for molecular identification to further determine the genetic relationship of the tested strains.The strain growth rate, biomass,mycelial length and other indicators under the condition of 23 ℃ (CK) and 30 ℃ high temperature stress were recorded. Result:All the 14 strains of Armillaria had the highest similarity and the closest relationship with Armillaria gallica,but there were significant differences in growth characteristics among different G. elata producing areas. The 14 strains of Armillaria were classified into Ⅳ groups,and the growth status was groupⅠ>group Ⅱ>group Ⅲ>group Ⅳ. After treatment with high temperature stress,the tolerance of each strain to high temperature also showed obvious differences,as shown in the average growth rate of the mycelial was GZ16>SX1>GZ1. The rank of relative mycelial length was GZ16>SX1>GZ3 and the relative biomass was GZ16>SX4>GZ1>HB1>AH2. Conclusion:Under high temperature stress,GZ16 was best in growth rate,relative length of mycelial,relative biomass and growth state,followed by SX1 and GZ1 strains. The results indicate that strains GZ16,SX1 and GZ1 have the strong resistance to high temperature and excellent growth characteristics at normal temperature,so these three strains are suitable to be produced in main G. elata producing areas in China.
摘要
Objective:Isolate and identify Mycena, expand the resources of geminating fungus of Gastrodia elata and optimize the culture conditions of Mycena,in order to provide information and guidance for the production of geminating fungus of G. elata. Method:Juvenile tuber tissue mass transfer separation and purification technology was used for the separation and purification of strains,traditional morphology microscopy was used to isolate the colony mycelia spores morphological characteristics, such as identification,polymerase chain reaction(PCR) amplification rDNA (Ribosomal DNA) internal transcribed spacer(ITS) was used for sequencing analysis and further homology with NCBI database retrieval,MEGA6 software was used to establish Phylogenetic tree by the Maximum likelihood method (MaximumLikelihood,M-L), so as to classify and identify isolated strains. At the same time,orthogonal test was used to optimize the optimal growth conditions of Mycena. Result:A total of 86 strains were isolated, which belong to 21 species in 12 genera. WMMFJ,SHXG,WMM-21 and MFJ8103 were identified as M. purpureofusca, and ZT01-6 and ZT01-8 were identified as M. cf. purpureofusca. The growth rate of Mycena in wheat bran medium was significantly higher than in PDA medium. The optimal medium composition for the growth of germinating bacteria was 100 g potato,150 g wheat bran and 20 g corn flour,100 g glucose. And 1,3,5-Trihydroxybenzene significantly promoted the growth of WMMFJ,and played a role in promoting the growth of WMM-21 and ZT01-6,and 2-Methoxyphenol promoted the growth of WMMFJ. Conclusion:Six strains of Mycena were isolated and identified,four of them are M. purpureofusca,and two of them are M. cf. Purpureofusca. The separation method improved the separation effect of germinating bacteria.
摘要
Gastrodia elata is a heterotrophic plant that needed to be symbiotic with Armillaria. The obstacle of continuous cropping in G. elata is serious during the G. elata cultivation, and the mechanism of obstacle in G. elata continuous cropping had not been solved. The planting of G. elata-Phallus impudicus is a new sequential planting pattern adopted in Guizhou province, but the effect of the cultivation on soil microbial community structure is still unclear. In this study, we collected four soil samples for the research including the soil without planted G. elata as control(CK), rhizosphere soil samples tightly adhering to the G. elata surface(GE), rhizosphere soil samples tightly adhering to Armillaria which was symbiotic with G. elata(AGE), the rhizosphere soil of P. impudicus planting after G. elata cultivation(PI). In order to explore the mechanism, the research study on the soil of G. elata-P. impudicus by using ITS and 16 S rDNA high-throughput sequencing technologies to detect soil microbial community structure including fungi and bacteria in the soil of CK, AGE, GE and PI. OTU clustering and PCA analysis of soil samples showed that the soil microbial diversity was relatively similar in AGE and GE. And the soil microbial in PI and CK clustered together. The results showed that AGE and GE had similar soil microbial diversity, as well as PI and CK. Compared with CK, the soil microbial diversity and abundance in AGE and GE were significantly increased. But the microbial diversity and abundance decreased in PI compared with AGE and GE. The annotation indicated that the abundance of Basidiomycota, Acidobacteria and Chloroflexi decreased, and that of Ascomycota, Zygomycota and Proteobacteria increased in AGE and GE compared with CK. In contrast to AGE and GE, PI was the opposite. The abundance of Basidiomycota, Acidobacteria and Chloroflexi increased in PI compared with AGE and GE. The abundance of microorganisms in the soil of PI and CK was similar. In addition, the co-culture of Armillaria and P. impudicus indicated that P. impudicus had obvious antagonistic effects on the growth of Armillaria. Therefore, it is speculated that the mechanism of G. elata-P. impudicus planting pattern related to the change of soil microbial. And we supposed that P. impudicus might inhibit the growth of Armillaria and change the soil microbial community structure and the abundance of soil microbial. And the soil microbial community structure was restored to a state close to that of uncultivated G. elata. Thus, the structure of soil microbial community planting G. elata could be restored by P. impudicus planting.
Subject(s)
Agaricales/growth & development , Bacteria/classification , Fungi/classification , Gastrodia/microbiology , Microbiota , Rhizosphere , Soil Microbiology摘要
The phenomenon that waste of fungus-growing materials in the planting process of Gastrodia elata is very common. It has been proved by practice that the used fungus-growing materials planted with G. elata can be used to plant Phallus impudicus. But the mechanism is unclear. In this study, we compared the different infested-capacity of Armillaria gallica and Phallus impudicus by morphological anatomy of the used fungus-growing materials. We also compared the differences on the two fungi consumed the main contents of fungus-growing materials, cellulose, lignin and hemicellulose, by using nitric acid-95% ethanol method, sulfuric acid method and tetrabromide method respectively, so that to explore the mechanism of A. gallica and P. impudicus recycle the fungus-growing materials, and to provide scientific basis for recycling the used fungus-growing materials of G. elata. The results showed that A. gallica had a strong ability to invade some parts outside the vascular cambium, but it had a weak ability to invade some parts inside the vascular cambium, while P. impudicus had a strong ability to invade the same parts. The contents of lignin and cellulose, which from inside and outside the vascular cambium of fungus-growing materials were significantly different. In the parts of outside the vascular cambium of fungus-growing materials, A. gallica degraded more lignin and cellulose, while P. impudicus degraded more hemicellulose. In the parts of inside the vascular cambium of fungus-growing materials, A. gallica degraded more cellulose, while P. impudicus degraded more hemicellulose. The present results suggested that A. gallica and P. impudicus made differential utilization of the carbon source in the fungus-growing materials to realize that P. impudicus recycle the used fungus-growing materials of G. elata. A. gallica used lignin and cellulose as the main carbon source, while P. impudicus used hemicellulose as the main carbon source.
Subject(s)
Agaricales/growth & development , Armillaria/growth & development , Cellulose/metabolism , Lignin/metabolism , Polysaccharides/metabolism摘要
Fungal disease is an important factor restricting the healthy development of Gastrodia elata industry. The control of fungal disease in G. elata is an important issue in production. This paper makes a detailed investigation on the current situation of G. elata disease in China through statistics on the failure rate, rotten pit rate and occurrence rate of G. elata disease in the main producing areas of China. It was found that G. elata disease was mainly infected from the top bud and junction, causing the occurrence rate of disease was 6%-17%, and the yield decreased by 10%-30%. The 23 dominant fungi were isolated from 18 typical G. elata disease samples. Through identification of colony morphology, mycelium morphology, spore morphology and genetic characteristics, they were finally identified as 13 species, belonging to 7 families and 7 genera. Trichoderma harzianum, Ilyonectria sp. and Ilyonectria destructans are the most frequently separated. Their isolation frequency were 22.22%,16.67%,16.67% respectively. Ilyonectria sp. and I. destructans were the first time isolated from G. elata disease samples. They may be the main pathogens causing soil-borne diseases of G. elata. T. harzianum has certain potential as Gastrodia biocontrol bacteria. This study can provide a theoretical basis for the research and development of control technology of Gastrodia fungi disease.
Subject(s)
China , Fungi/pathogenicity , Gastrodia/microbiology , Plant Diseases/microbiology摘要
The study is aimed to create seed materials and dissect the molecular mechanism of sexual propagation of Gastrodia elata. In this research, thirteen characteristics of flowers, flower stem, fruits, seeds and embryo of G.elata f. glauca and G.elata f. elata after bolting at room temperature(RT) and constant temperature(CT, 22 ℃) were determined. It was found that the constant temperature condition could prolong the bolting duration of G.elata and increased the number of flowers, while the variety of G.elata only affected the bolting duration, but had no effect on the number of flowers, and the G.elata f. elata was more likely to bolting than the G.elata f. glauca. The variety of G.elata was the main factor affecting the time of dehiscent fruit of G.elata, the temperature was the main factor affecting the fruits number and fruits diameter, and the constant temperature was more conducive to the fruits shape of G.elata than the room temperature. There was no significant difference in seed phenotype of G.elata varieties, but the seed embryo of G.elata seeds cultivated at constant temperature was fuller than that of G.elata cultivated at room temperature, and temperature had less influence on the seed phenotype of G.elata. But it was interesting to find that temperature and varieties had greater influence on the seed embryo of G.elata, constant temperature cultivation was more conducive to the formation of seed embryo of G.elata, and more the seed embryo of G.elata f. elata was easier to form than the seed embryo of G.elata f. glauca. However, the development of seeds and embryos of G.elata was significantly affected, and the development of seeds and embryos of G.elata f. glauca was more sensitive to temperature than G.elata f. elata. The research suggested that it is advisable for G.elata to produce seed materials by bolting at constant temperature(22 ℃).