2023 Vol. 38, No. 7
Display Method:
2023, 38(7): 761-771.
doi: 10.19303/j.issn.1008-0384.2023.07.001
Abstract:
Objective Effects of tms5 mutation site and related genetic regulatory factors in rice on the critical sterility-inducing temperature (CSIT) of the plant were investigated. Method Eleven CRISPR/Cas9 sequentially numbered T501 through T511 were designed to target 6 exons of tms5 to create mutants of Nipponbare (NIP, Oryza sativa ssp. Geng) and Minghui 86 (MH86, O. sativa ssp. Xian). Pollen male fertility and seed setting rate of the mutants were monitored under long day and high temperature field conditions in early August in Fuzhou (NHT) or at 22, 24, and 28 ℃ in phytotrons to identify the CSIT. Result The CSIT of the NIP mutants were higher than 28 ℃, while that of the MH86 mutants between 22 ℃ and 28°C. The tms5-1 T501 mutant had a significantly higher CSIT than the genetically identical tms5-2 counterparts T502, but the other 9 tm5 and the tm5-2 mutants did not differ on it. Expressions of the 3 UbL40 in young panicles were lower in the tms5-1 than the tm5-2 mutants of either rice varieties. Conclusion It appeared that the CSIT of tms5 mutants was affected by the genetic factors as well as the mutation site as the tms5-1 mutant T501 was shown significantly differed from the others.
2023, 38(7): 772-782.
doi: 10.19303/j.issn.1008-0384.2023.07.002
Abstract:
Objective Effects of seed sowing time and ambient temperature at grain-filling on the agronomic traits and rice quality/flavonoid content of Ziliangyou 737 were analyzed for improved the cultivation. Method Seeds of Ziliangyou 737 were sowed at intervals to vary the growing period and harvest season for studying the effects of sowing time and atmosphere temperature at grain-filling stage on the agronomic traits of the plants and quality and flavonoid content of the rice. In 7, 10, 14, and 21 d after start of grain-filling as well as at grain maturity, data were collected for a statistical analysis on the rice crops sowed on various dates. Result Under a shorter growing season, the plant height declined; the spike length became longer, then shorter; the seed setting rate and yield reduced, then increased; and the effective accumulated temperature (EAT) decreased gradually. The ∑H (High temperature hours) and ∑Th (Thermal accumulated temperature hour) within 10 d after grain-filling significantly affected the seeding rate of the mature seeds, so did the TA (Average temperature) within 14 d after grain-filling. When TA was 31.645 ℃, the seeding rate reached the lowest level, but the effect was lessened with increasing ambient temperature as the grain matured. High ∑Td (Daily thermal accumulated temperature) at 21 d after initial filling resulted in higher rate of failed grain-filling which translated to a lower seeding rate. VB5, VB6, and VB2 were the vitamins in the greatest amounts found in the grains, but no significant correlation was found between vitamin contents and sowing time. Late sowing caused linear decline in grain amylose due to decreased ∑H, ∑Td, and ∑Th. High temperature affected the amylose content positively, but the effect was less at the late filling stage. Controlling TA at a level below 26.84 ℃ could help maintain the quality of the waxy rice crop sowed in late season in Fujian. The contents of anthocyanins and flavonoids correlated to the sowing time as shown by the color of seed hull that changed from reddish brown to dark black. From 14 d after grain-filling till maturity, ∑Td, TA, and EAT became extremely significant factors in the color change as a rising temperature mitigated anthocyanins and flavonoids. Conclusion Scheduling of Ziliangyou 737 sowing significantly affected the growth and grain quality of the waxy rice. By controlling a TA less than 26.84 ℃ and an EAT at 2336.69–2390℃·d during grain-filling, crop yield and rice quality could be maximized. In Fujian, sowing between June 15 and 25 would thus be desirable. For other localities, cultivation in areas at high altitude or those that provide low TA at grain-filling stage would be recommended for Ziliangyou 737.
2023, 38(7): 783-791.
doi: 10.19303/j.issn.1008-0384.2023.07.003
Abstract:
Objective This study was conducted to analyze the genotypes of yield and quality-related traits of the purple waxy special rice Ziliangyou 737 and its parents, and to provide molecular basis for the further application of this variety and its parents in breeding and production. Method Using 13 molecular markers of 10 important genes, the alleles of Ziliangyou 737 and its parents Ziliang392s and Fuhui 737 were analyzed in relation to yield and quality. Result In 7 gene loci, both Zizliangyou 737 and its parents carry the same allele genotype. The high-yield allele Ha-Gn1a was carried in the seat of Gn1a, the major gene of panicles number. The long grain allele MH-GS3 was carried in the GS3 locus of the major gene for grain weight and grain length. The alleles of purple pericarp were carried in the Pb locus of seed coat gene. All alleles with low amylose content in waxy gene wx locus were carried in glutinous rice. In addition, the same allelic genotype was carried in the starch branch enzyme genes SBE1 and SBE3 and the limit dextrin enzyme gene PUL gene locus. In three loci, parents carry different alleles. In the Hd3a seat, the Zi 392S, Fuhui 737 and Ziliangyou 737 carry Hd3aNip, hd3aKasa and Hd3aNip/hd3aKasa respectively. On the Hd1 seat, carry Hd1jap, Hd1ind and Hd1jap/Hd1ind respectively. In the large subunit gene AGPlar locus of pyrophosphorylase, there were type I, type II and hybrid type I/II, respectively. Conclusion The genetic analysis of important agronomic traits of Ziliangyou 737 provided molecular evidence for its high yield, high quality, good adaptability and seed coat color, and provided reference for further utilization of related favorable alleles
2023, 38(7): 792-799.
doi: 10.19303/j.issn.1008-0384.2023.07.004
Abstract:
Objective Floral fragrance and release of Iris at different stages were analyzed using GC-MS. Methods Volatiles in the flowers of Iris lactea var. chinensis (Fisch.) Koidz at budding, blooming, and fading stages were determined by HS-SPME-GC-MS. Result The analysis identified 69 distinct aromatics. The continuously released volatiles in the entire flowering period were mainly ketones, terpenes, and aldehydes. There were also alcohols, esters, as well as in smaller quantities of alkanes, benzenoids, and nitrogenous compounds. At different flowering stages, hexanal, cyclohexanone, 5-hepten-2-one, 6-methyl- and β−irone were detected in the buds, hexanal, (−)-β−pinene, and 5-hepten-2-one, 6-methyl- in the blooms, and (-)-β−pinene, 5-hepten-2-one, 6-methyl-, hexanal, phenylethyl alcohol, and cyclohexanone in the fading flowers. The OPLS-DA showed differentiations on heptane, 3-methyl-, 5-hepten-2-one, 6-methyl-, (−)-β−pinene, hexanoic acid, ethyl ester, α−pinene, 3-carene, linalool, phenethyl alcohol, decanoic acid, ethyl ester, and β−irone at these stages. The calculated odor activity values (OAVs) on pyrazine, 2-methoxy-3-(1-methylpropyl)-, hexanoic acid, ethyl ester, linalool, nonanal, hexanal, 2-nonenal, and (E)- indicated that these were the characteristic aromatic substances of I. lactea var. chinensis and that pyrazine and 2-methoxy-3-(1-methylpropyl)- significantly greater in quantity (P＜0.01). Conclusion For the first time, the chemical composition of the fragrance of I. lactea var. chinensis flowers was analyzed, and compounds identified in detail. The released volatiles differed significantly during the floral development. It was at the fading stage when the fragrance peaked with the major aromatic contributors including hexanal, 5-hepten-2-one, 6-methyl-, cyclohexanone, and phenethyl alcohol.
2023, 38(7): 800-808.
doi: 10.19303/j.issn.1008-0384.2023.07.005
Abstract:
Objective Bioinformatics and expression of trehalose-6-phosphate synthase (TPS) gene in chili peppers,CaTPS9, were studied to understand its role in response to abiotic stress during the plant growth. Method CaTPS9 was cloned from Qiangfeng 101 to analyze the physicochemical properties, protein structure, cis-regulatory element, and phylogenetic tree. Expressions of the gene in the pericarp and placenta of a commercial pepper as well as the young pericarp, ripened pericarp, young placenta, ripened placenta, leaves, roots, stems, and flowers of chili pepper plants under varied stresses including low temperature, 3-indoleacetic acid(IAA), abscisic acid(ABA), salicylic acid(SA), gibberellin A3(GA3), and methyl jasmonate(MeJA) were determined using qRT PCR. Result The full-length cDNA sequence of CaTPS9 was 2604 bp encoded 867 amino acids. Located in the cytoplasm, the protein contained two conserved domains, Glyco_transf_20 and Trehalose_PPase, with a molecular weight of 97.60 kDa, an instability index of 44.27, and a theoretical isoelectric point of 5.63. It was a hydrophilic protein free of transmembrane structure and signal peptide sequence with a structure consisting mainly of alpha helixes and random coils. Phylogenetically, CaTPS9 was close to the homologous genes in Nicotiana tabacum, Solanum lycopersicum and Solanum tuberosum. It had hormones response, stress response, and plant growth and development cis−elements and was expressed most highly in the leaves, lowly in the placentas of a commercially available pepper, and significantly by 12 h SA induction. Stress, such as low temperature, IAA, ABA, GA3, and MeJA, significantly inhibited the expression. Conclusion It appeared that CaTPS9 might regulate the response of chili pepper plants to various abiotic stresses through the trehalose biosynthetic pathway.
2023, 38(7): 809-816.
doi: 10.19303/j.issn.1008-0384.2023.07.006
Abstract:
Objective NHX6 of Hippophae rhamnoides was cloned and expression under drought determined to study the stress resistance mechanism. Method HrNHX6 was cloned, amplified, and sequenced for analysis by biological software. Expression of the gene under drought stress was determined. Result Located on the tonoplast, HrNHX6 had no signal peptide and contained 13 transmembrane helical regions with numerous types of modification sites, such as N-glycosylation, protein kinase phosphorylation lation, myristoylation, amidation, and sugar transporter signature. The rusults of qRT-PCR analysis showed that the expression level of HrNHX6 gene was significantly different between different tissues under draught stress, and the expression level of HrNHX6 gene was significantly different between different drought stress treatment. Conclusion The drought resistance of H. rhamnoides was closely related to HrNHX6 as shown by the structure and expression under drought stress of the gene. The important role indicated a significant venue for improving the draught resistance of the medicinal plant.
2023, 38(7): 817-823.
doi: 10.19303/j.issn.1008-0384.2023.07.007
Abstract:
Objective Growth and root physiology of tomato seedlings and water-dissipation of substrate under ebb-and-flow irrigation in tray cultivation were studied. Methods Growth and root physiology of Ouxiu 201 Tomato seedlings as well as dissipation of moisture in the substrate irrigated by either ebb-and-flow or sprinkling method were monitored in a greenhouse experimentation. Results Under the ebb-and-flow irrigation, the seedling tray weight correlated linearly with the greenhouse temperature at a correlation coefficient of −0.846 and with the RH at 0.920. The weight loss due to evaporation increased with rising temperature at a coefficient of 0.966 and RH at −0.853. In comparison to sprinkling, the ebb-and-flow irrigation resulted in higher fresh weight, dry weight, volume, and length of roots at 4-leaves-and-one-bud stage of the seedlings by 44.81%, 45.91%, 93.81%, and 9.41%, respectively, and significantly elevated antioxidant enzyme activity of the roots as well as the Pn, and Gs of the plant, but significantly lower MDA content. Conclusion In the tray cultivation of tomato seedlings, the gain and loss of water in the substrate significantly correlated with the ambient temperature and RH under the ebb-and-flow irrigation which enhanced the seedling growth and fostered a vital root system with robust photosynthesis.
2023, 38(7): 824-832.
doi: 10.19303/j.issn.1008-0384.2023.07.008
Abstract:
Objective Contents and changes on aromatics in flavor-type grapes in various stages of fruit development were studied. Method Grapes from Red Globe, Seedless Venus, and Muscat Hamburg varieties were collected for aromatic determination by the headspace solid phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS).Changes in the contents of major aromatics in 3 growth stages were monitored and analyzed for principal components. Result Methyl salicylate, dimethyl phthalate, dibutyl phthalate, damascenone, and phenylethyl alcohol were the principal aromatic compounds in the fruits of Seedless Venus; linalool, α-Terpineol, geraniol, and geranic acid flavoring compounds, in those of Muscat Hamburg; and (E)-2-hexenal and 1-hexanol, in those of Red Globe.The content of ester aromatics continued to increase along the process of fruit development; the C13 isoprenoids (e.g., damascenone) were the highest in quantity at young stage; the volatiles, e.g., phenylethanol, accumulated from veraison to maturation; the aromatic terpenoids from young to veraison stage; and the C6 compounds displayed no consistent pattern in the entire growth period. Conclusion The contents and accumulation of aromatics in the varieties of grapes differed significantly. For instance, the aroma of Seedless Venus grapes came mainly from esters, C13 isoprenoids, and other aromatic compounds, with methyl salicylate, dimethyl phthalate, dibutyl phthalate, damascenone, and phenylethyl alcohol being the major contributors.Whereas Muscat Hamburg grapes composed largely of terpenes including linalool, α-Terpineol, geraniol, and geranic acid, while Red Globe consisted of C6 compounds like (E)-2-hexenal and 1-hexanol.
2023, 38(7): 833-841.
doi: 10.19303/j.issn.1008-0384.2023.07.009
Abstract:
Objective Effects of differential expression genes (DEGs) of Sparassis latifolia cultured with varied nitrogen sources on substrate degradation were analyzed for efficient cultivation of the mushroom. Method Cultured on substrates of varied nitrogen sources, such as ammonium sulfate (As), fish peptone (Pep), aspartic acid (Asp), and glutamic acid (Glu), S. latifolia mycelium growth was observed. Transcriptomes of DEGs were determined. Result Of the various nitrogen sources, Asp and Glu significantly promoted the mycelial growth (P＜0.01). According to a GO enrichment analysis, the DEGs in the mushrooms were mainly related to the activities of oxidoreductase, iron ion transmembrane transport, and iron assimilation by reduction and transport. And the KEGG showed their involvement in the metabolisms of tryptophan, arachidonic acid, and nitrogen. The expressions of the ostreolysin A6, GroES-like protein, and 6-methylsalicylic acid decarboxylase genes were significantly altered by the presence of Asp and Glu (P＜0.01). Conclusion The use of Asp and Glu as a nitrogen source promoted the substrate utilization through the heightened oxidoreductase activity and iron metabolism in S. latifolia. Hence, the addition would facilitate an efficient mushroom cultivation.
2023, 38(7): 842-850.
doi: 10.19303/j.issn.1008-0384.2023.07.010
Abstract:
Objective SSR molecular markers were used to verify the authenticity of F1 hybrids of Gerbera jamesonii Bolus. [Methods] From Rodrigo, Savannah, and their progenies, 65 reciprocal F1 hybrids of gerbera, primers with clear bands, high repeatability, and free of identical allele between parents were selected. Based on the transcriptomes, applicable means of species authentication were investigated. Results Of the 65 selected pairs of SSR primers, 26 were polymorphic at a rate of 40%. No identical alleles between the parents were found on the g24, g32, g38, and g64 primers with clear and distinct bands. They all contained a band of each parent in their amplified positive and negative F1 progenies and could correctly distinguish true from false hybrids. Meanwhile, the primer g24, g04, g44, and g39 could amplify new gerbera varieties/strains and also contained a band of each parent. However, they were limited to positively identifying genuine hybrids of Minghuifendai, Minghuihongyan, Meifen, and Huancai. Conclusion The SSR molecular markers selected could satisfactorily be used to authenticate the F1 hybrids of G. jamesonii Bolus for plant selection and commercial identification.
2023, 38(7): 851-856.
doi: 10.19303/j.issn.1008-0384.2023.07.011
Abstract:
Objective A rapid, accurate qRT-PCR method for detecting bovine kobuvirus (BKoV) was established and clinically tested for application. Method A pair of specific primers and a probe were designed and synthesized according to the 3D gene sequence of BKoV published on GenBank for the establishment of a TaqMan fluorescence qRT-PCR assay. The reaction system of the analytic was optimized prior to verification on clinical specimens. Result The optimal upstream and downstream primers were 300 nmol·L−1 in concentration, and the applied probe 400 nmol·L−1. A linearity with a correlation coefficient of R2= 0.999 was achieved in the range of 1×101−1×108 copies·μL−1. The amplification efficiency reached 103%. The assay demonstrated a high specificity among the pathogens related to diarrhea in calves, a high sensitivity with the minimum detection limit of 1×101 copies·μL−1 on BKoV plasmid standard, in comparison to that of 1×102 copies·μL−1 by conventional RT-PCR, and a high repeatability with a coefficient of variation of less than 3% within and between groups. In 37 bovine fecal samples collected from different pastures in Inner Mongolia from March to May 2021, the assay showed a positive detection rate of 24.3%. The calculated viral loads using the standard curve had an average on the feces from diarrheal animals at 3.7×105 copies·μL−1 and at 8.65×103 copies·μL−1 on the specimens from healthy calves. Conclusion The newly developed TaqMan fluorescent qRT-PCR assay was specific, repeatable, and sensitive in detecting BKoV. It could be applied for clinic diagnosis and epidemiological investigation on the disease.
2023, 38(7): 857-863.
doi: 10.19303/j.issn.1008-0384.2023.07.012
Abstract:
Objective Acute toxicities of 6 aquaculture drugs on juvenile Opsariichthys bidens were determined for safe applications. Method Medicine toxicity accumulation coefficients (MACs) of 6 drugs commonly applied for aquaculture were determined on the fish in a tank at a controlled temperature of 25.8–28.3 ℃. Result The drugs at specific doses displayed varied degrees of safety that ranked as Meiting Ⅱ (2.222 mg·L−1)＞Chelunchongbike (0.790 mg·L−1)＞povidone iodine (0.659 mg·L−1)＞Baitexindilin (0.225 mg·L−1)＞Baitezhihuandi (0.214 mg·L−1)＞Baidianjing (0.006 mg·L−1). The MACs of these drugs on O. bidens were positive but differed in the toxic effect to the fish due to differentiated accumulation and degradation of as well as sensitivity of the fish to the individual drugs. Conclusion According to the classification standards, Meiting Ⅱ was determined to be low toxic, while Chelunchongbike, povidone iodine, Baitexindilin, and Baitezhihuandi, moderately toxic, and Baidianjing, highly toxic to O. bidens juveniles.
2023, 38(7): 864-872.
doi: 10.19303/j.issn.1008-0384.2023.07.013
Abstract:
Objective Effects of different types of saline-alkali soil that are selenium (Se)-rich on the absorption, transport, and accumulation of Se in rice grown on them were studied. Method Saline-alkali-tolerant Ningjing 45 seedlings were grown on soils in the Yellow River irrigation area of Ningxia province that were mildly saline (Treatment A), mildly alkaline (Treatment B), and moderate-severely alkaline (Treatment C). Se uptake, accumulation, and quality of the seedlings grown on the pots were determined. Result (1) The Se contents in the roots, stems, leaves, and grains of the rice plants ranked as Treatment C＞Treatment B＞Treatment A, and at maturity, roots＞stems＞leaves＞grains. The contents in mature grains were 0.17 mg·kg−1 under Treatment A, 0.217 mg·kg−1 under Treatment B, and 0.24 mg·kg−1 under Treatment C. They all met the enrichment standard set for rice; however, Treatment C produced a significantly higher level of the metal element in the plants than the other two treatments (P＜0.05). (2) The Se accumulation in rice peaked at heading stage but varied in the organs at maturity with the ranking of milled rice＞stems＞leaves＞roots＞glume. (3) In seedling to heading stage, Se transported basically from the roots to the stems and leaves, but in heading to maturity, mainly from the stems and leaves to the spikes. (4) The Se absorption and primary transport coefficients of the roots increased with increasing salinity and alkalinity in soil. Thus, the effect was the greatest under Treatment C, followed by Treatment B and Treatment A. In contrast, the secondary transfer coefficient decreased with the increases to reverse the ranking order. Conclusion It appeared that Se-enrichment for rice grown on a saline-alkali soil that had an ample supply of the metal element could be realized if the soil had a pH ranging between 8.41 and 9.51, salt content between 1.74 g·kg−1 and 3.62 g·kg−1, and Se content of 0.23 mg·kg−1. And, interestingly, moderate and heavy saline-alkali soils bolstered the absorption, transport, and accumulation of Se in rice, and therefore, the normally considered undesirable conditions would enhance, rather than hinder, the use of like land masses.
2023, 38(7): 873-881.
doi: 10.19303/j.issn.1008-0384.2023.07.014
Abstract:
Objective Endophytes in sugarcane were screened for drought resistance to serve as microbial and germplasm resources. Methods Drought-tolerant microbes were isolated from a collection of 329 sugarcane endophytes at the Sugarcane Research Institute, Yunnan Agricultural University. In a simulated experiment, target strains were screened under the artificial draught condition created by using 0-45% PEG6000. Potential candidates were identified based on the morphological, physiological, and biochemical characteristics as well as 16sDNA sequences of the microbes. Optimal culture conditions for the growth, nitrogen fixation ability, and IAA secretion of the identified isolates were determined. Results Two strains that exhibited significant drought-tolerance were identified. K5 was a Kocuria sp. with an accession number of OP737815, and ZM a Bacillus sp. with an accession number of OP740361. Withstood the stress exerted by 40% PEG6000 or 100 g·L−1 sodium chloride, they could grow stably at 50 ℃ in wide ranges of pHs and temperatures but optimally under an initial pH at 5.0 for K5 and 6.0 for ZM. ZM was stronger than K5 in fixing nitrogen. Within a limited timeframe, the IAA secretion of the two strains increased with prolonged culture. Conclusion Both K5 and ZM displayed significant drought-resistance with the abilities of nitrogen fixation and IAA secretion. They could be potential candidates as a microbial agent, and their carrier sugarcane a valuable germplasm.
2023, 38(7): 882-888.
doi: 10.19303/j.issn.1008-0384.2023.07.015
Abstract:
Objective Effects of water activity (aw) and temperature on the growth and toxicity of Aspergillus flavus on mature grains of tartary buckwheat were studied to determine the safety conditions for harvest, processing, transportation, and storage of the crop. Method Varied medium aw (i.e., 0.87, 0.90, 0.93, 0.95 and 0.99) were created by additions of glycerol to culture A. flavus MI18-S1 isolated from the infected grains for a 7-day test at 15, 21, 28, 35 or 42 ℃. Colony diameter on petri dish was measured, spore count calculated from the blood cell counter board, and aflatoxin content determined by HPLC. Result The fungus was found to produce aflatoxins B1 and B2, predominately B1, but not G1 or G2. The highest spore count and toxicity were generated at 28 ℃ and aw=0.93. When the temperature was above 28 ℃, the colony could grow better under high water activity. At a temperature either below 15℃ or above 42 ℃, the fungal growth and toxin-production slowed or ceased. Within the range between these temperatures, the aflatoxin content increased at first and followed by a decline. A multivariate linear regression analysis showed no significant correlation between the colony diameter and the toxin production. Instead, the amount of aflatoxin generated in the culture was mainly affected by the aw and temperature of the environment, and most critically, sporulation of the fungus. And, sporulation of the fungus was significantly affected by the aw, temperature of the environment, the amount of aflatoxin generated in the culture and the colony diameter. Conclusion The growth and aflatoxin production of A. flavus on buckwheat grains were at peak when stored at 28 ℃ on a medium with aw of 0.93. Therefore, in many aspects of the buckwheat production chain, especially during the storage stage, it is advisable to avoid these optimal conditions in order to control the spore and toxin production of buckwheat, ensuring food safety. This study can provide a theoretical basis and data support for the development of preventive and control measures against Aspergillus flavus contamination during buckwheat harvesting, processing, transportation, and storage.
