2023 Vol. 38, No. 5
Display Method:
2023, 38(5): 515-523.
doi: 10.19303/j.issn.1008-0384.2023.05.001
Abstract:
Objective A new variety of super-sweet corn characterized with the desirable properties of high-quality, high-yield, wide-adaptability, and strong resistance to lodging, diseases, and hot climate was bred. Methods A tropical yellow-kernel, super-sweet inbred Mintian T146 line of corn germplasm was crossbred with Mintian AS67, a temperate white-kernel, super-sweet corn inbred line newly developed by the Crop Research Institute at Fujian Academy of Agricultural Sciences. The new variety of corn was submitted to regional trials followed by test production held in two regions in the nation. Results Super-sweet Minshuangse No. 6 bearing both yellow and white kernels on a cob was successfully bred in the winter of 2015. From 2018 to 2021, it was certified at the southeast and southwest regional trails for fresh consumption corn cultivars by the National Science and Enterprise Consortium. At the southeast regional trial from 2018 to 2019, Minshuangse No. 6 delivered an average fresh ear yield of 13,587.8 kg·hm−2, which was 7.6% higher than that of the reference Yuetian No. 16 with a yield point rate of 80.9%. A similar yield of 13 505.3 kg·hm−2 was shown at the southwest regional trial that was 3.6% higher than that of Yuetian 16 with a yield point rate of 70%. On the resistance to diseases under natural infectious conditions in the field, Minshuangse No. 6 was judged to be moderately resistant to the small spot disease, moderately to highly resistant to the grain wilt, moderately to highly resistant to the nodulation smuddery, moderately resistant to the stem rot, and moderately resistant to susceptible to the large spot disease and the southern rust. By artificial inoculation, it was identified to be susceptible to the head smut, susceptible to moderately resistant to the small spot, susceptible to the nodular smut, susceptible to the sheath blight, and susceptible to moderately resistant to the southern rust. In terms of quality, the comprehensive scores it received at the southeast regional trial was 89.2 and at the southwest 86.8, as compared to 85.0 for the reference. The physical and chemical quality analyses conducted by the Agricultural College at Yangzhou University indicated that the average residue percentage, total soluble sugars, and reducing sugars of the corn kernels sampled from Minshuangse No. 6 at the regional trials were 11.8%, 26.7%, and 11.25%, respectively. They were all higher than those of Yuetan 16. The new variety was officially granted certificates under the National Jade No. 20200523 in 2020 and the National jade No. 20220577 in 2022 by the National Crop Variety Certification Committee. In the 2021 and 2022 thousand-mu production tests, Minshuangse No. 6 had a production yield at Huizhou, Guangdong of 17 982 kg·hm−2, representing an increase of 5.9% over control and an improved revenue of 7 014 yuan·hm−2. And it scored higher than the reference at 90.8 on sensory evaluation. The results of the trial held at Jianou, Fujian, Minshuangse No. 6 showed a yield of 18 889.5 kg·hm-2, which was 9.44% higher than that of reference and 7 173.6 yuan·hm−2 more on revenue. It received 89.7 on the sensory score, which was also higher than the reference. Conclusion Minshuangse No. 6 displayed superior quality, yield, adaptability, and resistance to lodging, diseases, and heat. Consequently, it was recommended for promotion in the regions. Furthermore, the successful heterosis demonstrated by this study confirmed the validity of the "temperate germplasm × tropical germplasm" mode for corn breeding in general.
2023, 38(5): 524-529.
doi: 10.19303/j.issn.1008-0384.2023.05.002
Abstract:
Objective Stable transgenic glyphosate-tolerant strains of maize were obtained by screening T0-T3 generations of the genetically modified plants for the presence of EPSPS after an artificial exposure to glyphosate in the field. Method The gene, EPSPS, responsible for a maize plant to resist the herbicide was transferred into the excellent inbred line Jing 92 by means of the pollen tube pathway method. Expression of the introduced gene in maize was verified by molecular detection after exposing to glyphosate in the field. Result Fifteen resistant plants from T0 generation of the transgenic maize plants were identified after a treatment of 200 mg·L−1 glyphosate, and 10 by PCR. The genetic transformation rate was 1.03%. The identified plants were further scrutinized to eliminate non-resistant-gene-carriers. The proportion of confirmed carriers among the plants increased from the T1 to the T3 generation. Out of 5 strains in T3 generation, K3 and K8 were confirmed to have positively integrated EPSPS and be hereditarily stable. A test strip functional analysis on the two strains further verified the presence of the target gene. Conclusion The K3 and K8 transgenic lines identified by this study could plausibly be used to breed glyphosate-tolerant maize plants for cultivation.
2023, 38(5): 530-536.
doi: 10.19303/j.issn.1008-0384.2023.05.003
Abstract:
Objective Effects of methods in tilling the field in the regions of cold climate in China on rice quality and yield as well as soil organic matters were investigated for improved cultivation. Method Four different methods of tillage after rice harvest in preparation for the following planting season including (A) turning ground soil over in autumn, (B) tilling ground in autumn and breaking soil clusters in following spring, (C) plowing ground and pulping spent straws into soil in autumn, (D) returning and pulping stubbles to ground in autumn were employed for the testing. Effects of these varied method on the quality and yield of rice grown on the lots and the organic matters in the soil (i.e., total carbon in humus, total carbon of humic substances, and carbons of humic acid, fulvic acid, and humins) were monitored for a single factor randomized block experiment. Results Of the tried tillage methods, (D) delivered the highest contents of humus-related carbons in soil on an increasing trend with time, as the indicators were higher in 2020 than in 2019. The rice yield under (C) was significantly higher by 1.45 % over (B), 2.23 % over (A) and 1.50 % over (D). There was no significant difference due to the varied tillage on the processed grain quality, such as appearance and amylose content. However, (D) seemed to be better than (C) in that regard. Conclusions Tilling by plowing ground and pulping spent straws into soil in autumn (C) benefit the rice production in the following season, which was, therefore, recommended for practicing in the cold regions, such as the northeastern part of China. In addition, since (D) brought about the greatest improvement on the content of organic matters in the field soil, an autumn turning and pulping ground soil with stubbles was deemed to bring about a far-reaching effect for sustainable ecology.
2023, 38(5): 537-544.
doi: 10.19303/j.issn.1008-0384.2023.05.004
Abstract:
Objective Cloning and expression of mannose-1-phosphate guanyltransferase (GMP) gene that regulates the polysaccharide synthesis in Anoectochilus roxburghii were studied. Method ArGMP cDNA and genome sequences were cloned, and bioinformatics analyzed using online software. Gene expression pattern was then determined by qRT-PCR. Result The ORF of ArGMP was 1 086 bp encoded 361 amino acids with a length of 1 760 bp that contained 3 introns. It has an accession number of OQ030271 in GenBank. A stable hydrophilic protein free of a transmembrane structure, it was closely related to those of Dendrobium officinale, Apostasia shenzhenica, and Phalaenopsis equestris. Expressed differently in different tissues, the highest level was found in the flowers. The expression varied under different temperatures that peaked at 25 ℃, decreased significantly after 3 h at 35 ℃, and was severely inhibited beyond that. However, it was not affected by the stress exerted by NaCl in different concentrations. Conclusion The cDNA and sequence of GMP gene from A. roburghii were successfully cloned to display a tissue-specific expression pattern that could be significantly affected by temperature but not salt stress.
2023, 38(5): 545-551.
doi: 10.19303/j.issn.1008-0384.2023.05.005
Abstract:
Objective Effect of LED photoculture on yield of levodopa in broad bean sprouts was studied. Method Broad bean sprouts were exposed to LED of varied intensity and cultured for varied time periods in single factor and L9 (34) orthogonal experiments. Levodopa generated in the sprouts under the conditions was determined for process optimization. Result Low LED light enhanced the accumulation of sprout dry matters. However, it was under high intensity the productivity of levodopa in the sprouts was significantly encouraged. The optimum conditions for a maximized levodopa production of 19.62 g·m−2 were found to include 9 h of 500 lx LED exposure in 6 d of cultivation. Conclusion The photoculture of broad bean sprouts under intense LED lights could significantly promote the production of the functional amino acid, levodopa.
2023, 38(5): 552-558.
doi: 10.19303/j.issn.1008-0384.2023.05.006
Abstract:
Objective Argonaute (AGO) proteins play the important roles in a wide varieties of plant biological processes. To explore the cis --acting elements of DlAGO4 and DlAGO6 promoters, the expression patterns of recombinant vector of DlAGO4 and DlAGO6 promoters under different hormones treatments in longan, DlAGO4 and DlAGO6 promoters in longan were cloned and analyzed. Method Promoters of DlAGO4 and DlAGO6 from Dimocarpus longan were cloned by PCR for a bioinformatic analysis. Fusion expression vectors of the full-length promoters and GUS gene were constructed. The Agrobacterium tumefaciens-mediated transformation was performed on protocorm of tobacco leaves to observe the transient expression of the genes, while promoter activity examined by GUS histochemical staining. Result The sequence of DlAGO4 promoter was 1 514 bp and the sequence of DlAGO6 promoter was 1 784 bp. Both had the core elements of TATA- and CAAT-box as well as ababolic acid (ABA), methyl jasmonate (MeJA), and photoresponsive elements. The DlAGO4 promoter also contained salicylic acid (SA), gibberellin response, and anaerobic response regulatory elements, while the DlAGO6 promoter had auxin and circadian rhythm regulatory elements. The promoters exhibited a GUS-driving activity that was weaker than CaMV35S. The relative expression of GUS of PDlAGO6::GUS recombinant tobacco leaf was significantly elevated by the treatment of MeJA and ABA, whereas that of PDlAGO4::GUS recombinant tobacco leaf by SA and MeJA. Conclusion The promoters of DlAGO4 and DlAGO6 from longan were successfully cloned. As hormone-induced promoters, they could activate the GUS expression and be involved in the somatic embryo development and hormone response of plants.
2023, 38(5): 559-565.
doi: 10.19303/j.issn.1008-0384.2023.05.007
Abstract:
Objective Effects of placing a sunlight-reflecting film on the ground on the coloration and quality of Kyoho grapes were studied. Method In a rain shelter at Xiaosong Town, Jian 'ou City, Fujian, 3-year-old Kyoho grape vines were cultivated. In between rows, a silver-white aluminized plastic film was placed on the ground starting from the initiation of fruit color change till maturity. Effects of the film application on color of the grapes on upper and lower parts of a bunch as well as the anthocyanin content, longitudinal and transverse longitudes, shape index, as well as the contents of soluble solids, sugars, and total acid of the fruits during the ripening process were monitored. Results The ground-covering reflective film hastened the grape color formation, evened the color development of individual fruits on a cluster, and significantly increased the anthocyanin content in the peel. The firmness of the grapes increased by more than 67.30%, while the individual fruit weight rose by more than 26.79%. A uniform typical oval or oblong Kyoho grape shape was obtained on the vines. Although a slight increase on the soluble solids in the fruits by the film application was observed, it was not significant. Conclusion Covering ground with the reflective film 30d prior to fruit maturation significantly facilitated Kyoho grape vines in achieving the desirable uniformity on fruit color development and shape formation in a rain shelter. In addition, the application significantly increased anthocyanin content in the peel that could materially intensify the color enhancing the visual and bioactivity quality of the grapes for marketing.
2023, 38(5): 566-573.
doi: 10.19303/j.issn.1008-0384.2023.05.008
Abstract:
Objective Cryopreservation technology was applied for the conservation of the ornamental Malus Red Splendor crabapple germplasm. Methods Sucrose concentration and time for pretreatment as well as conditions for the subsequent loading, PVS2, and unloading treatments on the shoot tips of Malus spp. were studied. Under the selected preservation conditions, additional improvement on the post-freezing survival of the shoots by application of antioxidants (i.e., CAT, AsA, and GSH) and PCD inhibitors (i.e., Eth and SNP) was explored. Results (1) The procedures employed for vitrification to preserve the shoots included cutting the histoponic seedlings into 4-5 mm segments containing terminal buds, inoculating the cut stems segments in a pre-culture medium containing 0.7 mol·L−1 sucrose at 4 ℃ for 2 d, cutting the tips into approximately 1.5-2 mm in length and placing in a loading solution for 20 m at room temperature, treating with PVS2 solution for 90 m at 0 ℃ followed by immediate submerging in liquid nitrogen, removing the cryogenically frozen tips to rapidly thaw them in a water bath at 38 ℃ for 1 m, and rinsing twice with an unloading solution at room temperature with constant shaking for 10 m each time. The post-cryopreservation survival rate of the shoot tips was 66.58%, and 16.67% of them recovered to grow as normal. (2) The best improvement by the addition of various agents was found to be CAT at 200U·mL−1, AsA at 600 μmol·L−1, and GSH at 0.04 μmol·L−1, which increased the post-freezing survival rate by 20.28%, 6.75%, and 27.61%, respectively, over control. On the other hand, the addition of Eth or SNP posted no significant effect on the tip survival. Conclusion The survival rate of the post-cryopreservation Malus Red Splendor shoot tips could be further improved by the addition of GSH, CAT or AsA in appropriate concentration. GSH could most significantly raise the rate of viable plants after the cryopreservation from 16.67% to 41.39%.
2023, 38(5): 574-582.
doi: 10.19303/j.issn.1008-0384.2023.05.009
Abstract:
Objective Recombinant protein of chalcone isomerase (CHI) gene in Rhododendron delavayi was prepared and its activity was verified. This will provide theoretical basis for analyzing the function of CHI, and also lays a foundation for improving flower color and increasing medicinal ingredientst. Method Primers were designed based on the sequence of RdCHI1, and the prokaryotic expression vector constructed. Conditions to induce the expression of soluble recombinant protein were optimized. Activity of the prepared protein was verified by an in vitro enzymatic assay. Result The successfully constructed RdCHI1 prokaryotic expression vector was expressed in the supernatant under the optimized induction that applied 0.35 mmol·L−1 IPTG at 15 ℃ for 36 h. The nickel column-purified recombinant protein significantly hastened the conversion of naringin chalcone to naringin in an in vitro assay. Conclusion As a type I CHI, RdCHI1 significantly accelerated the convertion from naringin chalcone to naringin and increase the accumulation od flavonoids.
2023, 38(5): 583-597.
doi: 10.19303/j.issn.1008-0384.2023.05.010
Abstract:
Objective FtsH gene plays an important role in plant stress resistance. Revealing the function and expression pattern of FtsH gene in tea plant can provide a theoretical basis for the improvement of tea plant resistance to photooxidation and high temperature stress. Method The FtsH gene was identified by tea plant genome data, and a CsFtsH31 gene sensitive to high temperature was screened by bioinformatics analysis. The CsFtsH31 gene was overexpressed in tobacco, and the expression patterns of CsFtsH31 gene against photooxidation and high temperature were analyzed. Result A total of 45 FtsH genes were identified in the genome of tea plant, which were divided into five categories according to the evolutionary relationship. The members of this family are distributed on different chromosomes of tea plants. CsFtsH genes in tea plants are differentially expressed in different tissues and stress conditions. After high temperature treatment, CsFtsH14, CsFtsH31 and CsFtsH34 genes in tea plants showed an increasing trend over time, and CsFtsH31 is particularly sensitive to high temperature. The transgenic tobacco with CsFtsH31 gene was obtained by genetic transformation of CsFtsH31 gene. The transgenic tobacco with CsFtsH31 gene was subjected to stress treatment. The results showed that the chlorophyll and soluble sugar content and superoxide dismutase activity of transgenic tobacco were higher than wild type, while malondialdehyde was lower than wild type. Conclusion The CsFtsH31 gene was expressed in tobacco by increasing the content of chlorophyll and soluble sugar, and the activity of superoxide dismutase, reducing the content of malondialdehyde, scavenging the reactive oxygen species produced by stress, protecting the function of membrane structure, and improving the resistance of tobacco to light oxidation and high temperature stress.
2023, 38(5): 598-606.
doi: 10.19303/j.issn.1008-0384.2023.05.011
Abstract:
Objective Physiological functions of the Sec pathway subunit secD of Aeromonas hydrophila were studied. Method An secD-knockout A.hydrophila was constructed by homologous recombination with pRE112 suicide vector. Effects of the genetic deletion on the physiological phenotypes including hemolytic and extracellular protease activities were determined on sheep’s blood and skim milk agar plates; on the biofilm formation, by crystal violet staining combined with SpectraMax® i3; on the bacterial acid-base and hyperosmolarity tolerance, according to the growth curve; and on the minimal inhibitory concentration (MIC) of antibiotics, using the broth microdilution method. Result Compared to wild-type, the constructed ∆secD A.hydrophila showed no significant differences in growth and tolerance to pH 5, 6, 7, and 8. The activities of hemolytic and extracellular protease, as well as the resistance to high osmotic pressure or pH 9 were enhanced. The biofilm forming ability significantly declined. And the resistance to antibiotics varied, as the MIC increased 4 times on oxytetracycline, tetracycline, enoxacin, or meropenem and two times on ciprofloxacin and norfloxacin, but decreased 2 times on erythromycin and 4 times on cefotaxime. Conclusion The Sec pathway subunit secD was confirmed to be involved in the transport of virulence factors and resistance to antibiotics in A.hydrophila. It could be a key element in studies for developing antimicrobial applications and prevention of disease infected by the pathogen.
2023, 38(5): 607-615.
doi: 10.19303/j.issn.1008-0384.2023.05.012
Abstract:
Objective Mitochondrial genome and phylogenetic relationship in Staphylininae subfamily of Philonthus spinipes (Sharp, 1874) were determined. Method The high-throughput sequencing technique was applied to secure the full length mitochondrial genome of P. spinipes. A phylogenetic analysis was conducted to decipher the beetle’s relationship in the Staphylininae subfamily. Using 21 exemplars of Staphylininae and 8 of Paederinae as ingroups as well as two Pselaphinae species as outgroups, the phylogeny was reconstructed with the maximum likelihood (ML) and Bayesian inference (BI) methods. Result The mitochondrial genome of P. spinipes was a circular molecule of 16 219 bp with the GenBank accession number of OL998729. It contained 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a non-coding control region. Most of the protein-coding genes started with ATT, ATA, or ATG, but nad6 did with ATC and nad1 with TTG. And cox1, cox2, cox3, and nad5 had an incomplete T or TA as the stop codon, while the remaining 9 genes terminated with TAA or TAG. All tRNA genes, except trnS1 which lacked the DHU arm, could be folded into a typical cloverleaf-like structure. The lengths of rrnL and rrnS were 1 275 bp and 765 bp with the A+T contents of 79.84% and 76.47%, respectively. Conclusion The two applied phylogenetic inference methods produced a similar tree topological structure that showed Paederinae to be monophyletic, Staphylininae non-monophyletic, and Paederinae nested within the Staphylininae subfamily.
2023, 38(5): 616-623.
doi: 10.19303/j.issn.1008-0384.2023.05.013
Abstract:
Objective Full-length cDNA of Caffeoyl-CoA-O-methyltransferase gene (CCoAOMT) in the lignin metabolism pathway of soybean was cloned, and bioinformatics analyzed to study the resistance mechanism to cyst nematode (Heterodera glycines Ichinohe) of the plant. Method CCoAOMT sequence from roots of a cyst nematode-resistant Huipizhiheidou soybean plant was cloned by RT-PCR. Results The full length cDNA of GmCCoAOMT was cloned and then submitted to GenBank with the accession number of MW480860. The length of GmCCoAOMT was 848 bp, containing an ORF of 741bp and encoding 246 amino acids with a predicted molecular weight of 27.6 kDa and a theoretical isoelectric point of 5.67. It was located in the cytoplasm with a conserved AdoMet_MTases domain and belonged to that superfamily. The multiple alignment on amino acid sequence and phylogenetic tree indicated a high similarity between the gene and those of other leguminous plants. Conclusion The cDNA of GmCCoAOMT was successfully cloned and shown with a close evolutionary relationship with Vigna unguiculata (Linn.) Walp., Cajanus cajan (Linn.) Millsp., Phaseolus vulgaris Linn. and Vigna angularis (Willd.) Ohwi et Ohashi but far from Lupinus albus and Cicer arietinum Linn.
2023, 38(5): 624-631.
doi: 10.19303/j.issn.1008-0384.2023.05.014
Abstract:
Objective EndogenousPseudomonas sp. in the root nodules of Hippophae thibetana that displayed a biological activity were isolated for toxicological identification and growth-promoting effect determination for the development of a biological agent for crop fertilization. Methods Strains of Pseudomonas sp. were isolated from the root nodules of H. thibetana by pure culture method. They were identified by morphology, physiological biochemistry, and 16S rDNA sequence, as well as examined for the growth-promoting effect on the host plant by artificial inoculation for evidence of phosphorus-dissolving and IAA-, iron- and cellulase-producing capacities. Results Out of the isolated endophytic strains, 4 showed greater than 99% 16S rDNA alignment with the reference Pseudomonas. Among them, QY-X10 and QY-X22 produced 400 mg·L−1 of solubilized phosphorus; QY-X4 yielded the highest IAA at 1.9 mg·L−1; all 4 strains contained iron-generating carriers; except QY-X10, all could degrade cellulose; QY-X6 promoted seed germination; and QY-X6 and QY-X10 helped the host plant to grow significantly more leaves and longer seedlings, while QY-X10 and QY-X22 did significantly to make the maximum-length leaves longer than did control. Conclusion There were 4 Pseudomonas sp. identified in this study that exhibited significant abilities to dissolve inorganic phosphorus and produce IAA and iron. Three of them could also degrade cellulose. By inoculating these strains on the host plants, the seed germination and seedling growth were effectively promoted. It appeared that they could be applied for crop fertilization.
2023, 38(5): 632-638.
doi: 10.19303/j.issn.1008-0384.2023.05.015
Abstract:
Objective Saccharomyces cerevisiae JP2, isolated from naturally fermented loquat, is a yeast strain with good metabolism of malic acidwine mash. To further investigate its intrinsic acid lowering mechanism, whole genome sequencing and genome sequence information analysis were conducted, and possible metabolic pathways of malic acid were analyzed to identify genes that affect acid lowering. Method Based on the Illumina Miseq and PacBio sequencing platforms, second and third generation sequencing techniques were used to sequence the entire genome of S. cerevisiae JP2 and the type strain S. cerevisiae S288c. Bioinformatics methods were used to assemble their sequences, predict their genes, and annotate their functions, while exploring the possible malic acid metabolism pathways of JP2. Result (1) A total of 52 contigs were obtained by assembling the genome of S. cerevisiae JP2, with a total genome size of 11.98 Mbp and a GC content of 38.07%; There is no significant difference in overall genome size and GC content between JP2 and S288c. (2) There are three pathways for JP2 to metabolize malic acid: malic acid → oxaloacetate → pyruvate; malic acid → pyruvate; and malic acid → fumaric acid. The main genes involved are maeA, MDH, and fumC, among which malic acid → oxaloacetate → pyruvate is the main metabolic pathway with the key gene maeA. (3) The simulated fermentation process showed a decrease of 35.8% in malic acid metabolism and key gene expression levels in JP2, with a significant decrease compared to S288c; Compared with S288c, the maeA gene of JP2 was significantly upregulated (P＜0.01), and its relative expression reached 130% of the initial value in the late fermentation stage. Conclusion JP2 exhibits high expression of maeA gene during fruit wine fermentation, significantly improving the efficiency of malic acid degradation. The research results can provide technical support for the study of the acid reducing functional gene of S. cerevisiae JP2 and the development of acid reducing engineering bacteria.
