OsPPG encodes a pseudouridine-5'-phosphate glycosidase and regulates rice spikelet development

Florets are the basic structural units of spikelets,and their morphogenesis determines the yield and quality of rice grains.However,whether and how pseudouridine-5’-phosphate glycosidase participates in rice spikelet development remains an open question.In this study,we identified a novel gene,OsPPG,which encodes a peroxisome-localized pseudouridine-5’-phosphate glycosidase and regulates the development of rice spikelets.osppg mutants exhibited abnormal sterile lemma,lemma,palea,lodicule,stamens,and pistils;male sterility;shorter panicles;and reduced plant height.OsPPG was found to regulate several OsMADS genes,thereby affecting the morphogenesis of rice spikelets.Furthermore,metabolomics revealed that the OsPPG gene was involved in the decomposition of pseudouridine via the pyrimidine metabolism pathway and may affect the jasmonic acid signaling pathway.These results suggest that OsPPG is a key regulator of rice spikelet development.

Integration of 1H-NMR-Based Metabolomics Approach with Traditional Chinese Pulse Diagnosis to Evaluate the Qi-Enhancing Activity of Herbal Tea

Human metabolism is intricately linked to an individual’s health status. Regardless of living habits, it will be reflected in the metabolic characteristics of urine. The utilization of the 1H NMR-based metabolomics method has enabled examine the metabolomic changes in urine under various physiology conditions, providing valuable insights into metabolites. In this particular study, volunteers were divided into two groups based on the strength of their spleen pulses, using the pulse diagnosis method employed in traditional Chinese medicine. Subsequently, their urine samples were analyzed, revealing notable variances in urea, creatinine, citric acid, succinic acid, trimethylamine-N-oxide (TMAO), alanine, hippuric acid, and glycine between the two groups. Interestingly, individuals with weak spleen pulses showed significant improvements after consuming herbal tea. Furthermore, we conducted LC-MS analysis on herbal tea and performed adenosine triphosphate (ATP) activity tests on the C2C12 mouse skeletal muscle cell line. The results indicated that within a reasonable concentration range, exposure to herbal tea led to an increase in the mitochondrial ATP production capacity of C2C12 cells. These findings shed light on the relationship between traditional Chinese medicine pulse diagnosis and urine metabolites, highlighting their potential as non-invasive and straightforward health assessment indicators. They can aid in the preliminary determination of necessary dietary and lifestyle changes to enhance overall bodily health.

Root Endophyte Shift and Key Genera Discovery in Rice under Barnyardgrass Stress

Despite increasing knowledge of barnyardgrass(Echinochloa crus-galli) interference with rice, relatively little is known how endophytes improve the ability of rice against barnyardgrass stress. Here, we provided a detailed temporal characterization of rice root-associated microbiomes during co-cultivation with barnyardgrass and a comparison with the microbiomes of weed-free rice plants. Alpha diversity analysis indicated that barnyardgrass had the opposite effects on endophytic bacteria and fungi in rice roots, in terms of the community diversity, richness and coverage at the rice seedling stage. Principal coordinate analysis showed that barnyardgrass had only a minor effect on the community composition of endophytes in rice roots at the rice seedling stage, but showed a significant and maximum interference at the heading stage. Rice recruited many endophytes to resist biotic stress from barnyardgrass, especially for fungi. PICRUSt(phylogenetic investigation of communities by reconstruction of unobserved states) predictive analysis indicated that 23 metabolic pathways of bacteria were overrepresented in rice. In addition, the main trophic mode of fungi was pathotroph according to FUNGuild analysis. A positive correlation between bacteria and fungi in rice roots was found via network analysis. Anaeromyxobacter, Azospira and Pseudolabrys were the vital bacteria, Phaeosphaeria and Funneliformis were the dominant fungi in maintaining the stability of the ecological network. These results provided data and a theoretical basis for the in-depth understanding of what role endophytes play in rice resistance to barnyardgrass stress and will have implications on improving the resistance of rice against biotic stress using root microbiota.

High-throughput discovery of plastid genes causing albino phenotypes in ornamental chimeric plants

Chimeric plants composed of green and albino tissues have great ornamental value.To unveil the functional genes responsible for albino phenotypes in chimeric plants,we inspected the complete plastid genomes(plastomes)in green and albino leaf tissues from 23 ornamental chimeric plants belonging to 20 species,including monocots,dicots,and gymnosperms.In nine chimeric plants,plastomes were identical between green and albino tissues.Meanwhile,another 14 chimeric plants were heteroplasmic,showing a mutation between green and albino tissues.We identified 14 different point mutations in eight functional plastid genes related to plastid-encoded RNA polymerase(rpo)or photosystems which caused albinism in the chimeric plants.Among them,12 were deleterious mutations in the target genes,in which early termination appeared due to small deletion-mediated frameshift or single nucleotide substitution.Another was single nucleotide substitution in an intron of the ycf3 and the other was a missense mutation in coding region of the rpoC2 gene.We inspected chlorophyll structure,protein functional model of the rpoC2,and expression levels of the related genes in green and albino tissues of Reynoutria japonica.A single amino acid change,histidine-to-proline substitution,in the rpoC2 protein may destabilize the peripheral helix of plastid-encoded RNA polymerase,impairing the biosynthesis of the photosynthesis system in the albino tissue of R.japonica chimera plant.

Enhanced removal of nitrate and phosphate from wastewater by Chlorella vulgaris: Multi-objective optimization and CFD simulation

To enhance the efficiency of wastewater biotreatment with microalgae, the effects of physical parameters need to be investigated and optimized. In this regard, the individual and interactive effects of temperature, p H and aeration rate on the performance of biological removal of nitrate and phosphate by Chlorella vulgaris were studied by response surface methodology(RSM). Furthermore, a multi-objective optimization technique was applied to the response equations to simultaneously find optimal combinations of input parameters capable of removing the highest possible amount of nitrate and phosphate. The optimal calculated values were temperature of 26.3 °C, pH of 8 and aeration rate of 4.7 L·min^(-1). Interestingly, under the optimum condition, approximately 85% of total nitrate and 77% of whole phosphate were removed after 48 h and 24 h, respectively, which were in excellent agreement with the predicted values. Finally, the effect of baffle on mixing performance and, as a result, on bioremoval efficiency was investigated in Stirred Tank Photobioreactor(STP) by means of Computational Fluid Dynamics(CFD). Flow behavior indicated substantial enhancement in mixing performance when the baffle was inserted into the tank. Obtained simulation results were validated experimentally. Under the optimum condition, due to proper mixing in baffled STP, nitrate and phosphate removal increased up to 93% and 86%,respectively, compared to unbaffled one.

Quinclorac Resistance in Echinochloa crus-galli from China

Echinochloa crus-galli is a major weed in rice fields in China,and quinclorac has been long used for its control.Over-reliance of quinclorac has resulted in quinclorac resistance in E.crus-galli.Two resistant(R)E.crus-galli populations from Hunan,China were confirmed to be at least 78-fold more resistant to quinclorac than the susceptible(S)population.No difference in foliar uptake of 14C-labelled quinclorac was detected between the R and S plants.However,a higher level of 14C translocation and a lower level of quinclorac metabolism were found in the R plants.Basal and induced expression levels ofβ-cyanoalanine synthase(β-CAS)gene andβ-CAS activity were not significantly different between the R and S plants.However,the induction expression of 1-aminocyclopropane-1-carboxylic acid oxidase(ACO1)gene by quinclorac treatment was evident in the S plants but not in the R plants.Quinclorac resistance in the two resistant E.crus-galli populations was not likely to be related to foliar uptake,translocation or metabolism of quinclorac,nor to cyanide detoxification viaβ-CAS.Thus,target-site based quinclorac signal reception and transduction and regulation of the ethylene synthesis pathway should be the focus for further research.

The power of microsatellite markers and AFLPs in revealing the genetic diversity of Hashemi aromatic rice from Iran

Hashemi, a popular aromatic rice among Iranians, is famous for its fragrance and taste. Such features are major reasons for its higher price compared to non-aromatic varieties available in Iran. Therefore, the knowledge of genetic diversity of this profitable crop is a fundamental ineterst for plant breeders in future breeding programs. In the present research, genetic diversity among 35 genotypes of Hashemi aromatic rice （Oryza sativa L.） from Guilan and Mazandaran provinces of Iran was estimated using simple sequence repeat （SSR） and amplified fragment length polymorphism （AFLP） markers. Out of 21 SSR and 12 EcoRI-Msel AFLP marker combinations, only 16 SSRs and 10AFLPs exhibited polymorphic patterns while others were monomorphic. The 10 AFLP primer combinations produced a total of 142 of bands and 20 were polymorphic （14.08%）. Moreover, 40 out of 47 bands amplified with 16 SSR markers showed polymorphism （85.1%）. The average num- ber of alleles identified by SSR primers was 2.56 alleles per locus with a range of 2 to 4. The average value of polymorphic information content （PIC） was 0.393 and 0.468 for AFLP and SSR markers, respectively. However, the genetic similarity values ranged from 0.26 to 1 for SSRs and 0.21 to 1 forAFLPs. Later, a unweighted pair group method with arithmetic mean （UPGMA） dendrogram was generated and genotypes were clustered into four groups with SSRs at similarity coefficient of 0.55 while AFLPs clustered them into six groups at similarity coefficient of 0.41. Cluster analysis revealed a narrow genetic diversity and low correlation between geographical differentiation and genetic distance within cultivars.

Primordial germ cell-mediated transgenesis and genome editing in birds

Transgenesis and genome editing in birds are based on a unique germline transmission system using primordial germ cells(PGCs), which is quite different from the mammalian transgenic and genome editing system. PGCs are progenitor cells of gametes that can deliver genetic information to the next generation. Since avian PGCs were first discovered in nineteenth century, there have been numerous efforts to reveal their origin, specification, and unique migration pattern, and to improve germline transmission efficiency. Recent advances in the isolation and in vitro culture of avian PGCs with genetic manipulation and genome editing tools enable the development of valuable avian models that were unavailable before. However, many challenges remain in the production of transgenic and genome-edited birds,including the precise control of germline transmission, introduction of exogenous genes, and genome editing in PGCs.Therefore, establishing reliable germline-competent PGCs and applying precise genome editing systems are critical current issues in the production of avian models. Here, we introduce a historical overview of avian PGCs and their application, including improved techniques and methodologies in the production of transgenic and genome-edited birds, and we discuss the future potential applications of transgenic and genome-edited birds to provide opportunities and benefits for humans.

Effect of post-harvest UV-C irradiation and calcium chloride on enzymatic activity and decay of tomato(Lycopersicon esculentum L.)fruit during storage

Tomato is one of the extensively consumed vegetable crops worldwide. The regular consumption of tomato decreases the incidence of chronic degenerative diseases such as certain types of cancer and cardiovascular diseases. The objective of this study was to find an appropriate method that not only reduces tomatoes decay, but also maintains its post-harvest quality. A factorial experiment based on randomized complete block design with three replications was conducted to evaluate effects of ultraviolet（UV）-C and CaCl_2 applications on tomato during storage. The traits studied included ethylene, polygalacturonase（PG） activity, pectin methyl esterase（PME） activity, firmness, total phenol content, and fungal-induced decay were measured weekly during 35 days of storage. Both UV-C and CaCl_2 treatments had positive effects on tomato quality as compared to control treatment. The 3 and 4.5 k J m^（-2） levels of UV-C and 2% CaCl_2 had positive effects on quality characteristics, respectivly. Fruits treated by UV-C and CaCl_2 had higher phenol and firmness, and less PME activity, PG activity, ethylene production, and decay than the control fruits. In conclusion, increasing in storage duration significantly affected the fruits quality by increasing in ethylene, PME activity, PG activity, decay and decreasing the phenol content and firmness. But UV-C and CaCl_2 led to significant decrease in this adverse impact relative to control treatment.

Acquisition of pluripotency in the chick embryo occurs during intrauterine embryonic development via a unique transcriptional network

Background: Acquisition of pluripotency by transcriptional regulatory factors is an initial developmental event that is required for regulation of cell fate and lineage specification during early embryonic development. The evolutionarily conserved core transcriptional factors regulating the pluripotency network in fishes, amphibians, and mammals have been elucidated. There are also species-specific maternally inherited transcriptional factors and their intricate transcriptional networks important in the acquisition of pluripotency. In avian species, however, the core transcriptional network that governs the acquisition of pluripotency during early embryonic development is not well understood.Results: We found that chicken NANOG(c NANOG) was expressed in the stages between the pre-ovulatory follicle and oocyte and was continuously detected in Eyal-Giladi and Kochav stage I(EGK.I) to X. However, c POUV was not expressed during fol iculogenesis, but began to be detectable between EGK.V and VI. Unexpectedly, c SOX2 could not be detected during fol iculogenesis and intrauterine embryonic development. Instead of c SOX2, c SOX3 was maternally inherited and continuously expressed during chicken intrauterine development. In addition, we found that the pluripotency-related genes such as c ENS-1, c KIT, c LIN28 A, c MYC, c PRDM14, and c SALL4 began to be dramatical y upregulated between EGK.VI and VII.Conclusion: These results suggest that chickens have a unique pluripotent circuitry since maternally inherited c NANOG and c SOX3 may play an important role in the initial acquisition of pluripotency. Moreover, the acquisition of pluripotency in chicken embryos occurs at around EGK.VI to VI I.

Validation of EST-derived STS markers localized on Qfhs.ndsu-3BS for Fusarium head blight resistance in wheat using a‘Wangshuibai’derived population

A few EST-derived STS markers localized on Qfhs.ndsu-3BS, a major QTL for resistance to Fusarium head blight （FHB） in wheat, have been previously identified in the ＇Sumai 3＇/＇Stoa＇ population. In this study, we used a ＇Wangshuibai＇ （resistant）/＇Seri82＇ （susceptible） derived population, linkage group, QTL, and quantitative gene expression analysis to assess the genetic background dependence and stability of the EST-derived STS markers for use in marker aided selection to improve FHB resistance in wheat. Based on our results, a QTL in the map interval of Xsts3B-138_1-Xgwrn493 on chromosome 3BS was detected for FHB resistance, which accounted for up to 16% of the phenotypic variation. BLASTN analysis indicated that Xsts3B-138_1 sequence had significant similarity with the resistance gene analogue. Real-time quantitative PCR showed that the relative expression of Xsts3B-138_1 in ＇Wangshuibai＇ at 96 h after inoculation was 2.6 times higher than ＇Seri82＇. Our results underlined that EST-derived STS3B-138 markers could be predominantly used in marker aided selection to improve FHB resistance in wheat.

Progress in drug delivery system for fibrosis therapy

Fibrosis is a necessary process in the progression of chronic disease to cirrhosis or even cancer,which is a serious disease threatening human health.Recent studies have shown that the early treatment of fibrosis is turning point and particularly important.Therefore,how to reverse fibrosis has become the focus and research hotspot in recent years.So far,the considerable progress has been made in the development of effective anti-fibrosis drugs and targeted drug delivery.Moreover,the existing research results will lay the foundation for more breakthrough delivery systems to achieve better anti-fibrosis effects.Herein,this review summaries anti-fibrosis delivery systems focused on three major organ fibrotic diseases such as liver,pulmonary,and renal fibrosis accompanied by the elaboration of relevant pathological mechanisms,which will provide inspiration and guidance for the design of fibrosis drugs and therapeutic systems in the future.

Analysis of Rhizosphere Microbial Community Structure in Different Pathogenesis Stages of Watermelon Fusarium Wilt

High-throughput sequencing technique was applied to analyze the microbial community structure of rhizosphere soil at different stages of watermelon fusarium wilt to find out the difference of dominant microbial community in rhizosphere during the occurrence of watermelon fusarium wilt.Illumina-Hiseq high-throughput sequencing platform was used to sequence 16S and ITS rDNA in rhizosphere soil.The soil was named CK1 before planting,CK2 at peak stage and CK3 at stable stage.The results showed that the soil bacterial diversity was in the order of CK1>CK3>CK2,indicating no significant difference between CK1 and CK3 and a significant difference between CK2 and CK1,CK3.At the genus level,the dominant bacteria were Mizugakiibacter(21.9299%),Rhodanobacter(5.0933%),and Lactobacillus(3.1921%).The diversity of soil fungi was in the order of CK1>CK3>CK2,all showing significant differences.At the genus level,the dominant fungus was Lysurus(54.4601%),Papulaspora(12.4252%),Acrophialophora(3.1729%).The results showed that the diversity and abundance of bacteria and fungi in rhizosphere soil decreased during the peak period of watermelon fusarium wilt.With the gradual stabilization of the disease,the diversity and abundance of bacteria and fungi in rhizosphere soil recovered to a certain extent.

Two novel gene-specific markers at the Pik locus facilitate the application of rice blast resistant alleles in breeding

Blast,a disease caused by Magnaporthe oryzae,is a major constraint for rice production worldwide.Introgression of durable blast resistance genes into high-yielding rice cultivars has been considered a priority to control the disease.The blast resistance Pik locus,located on chromosome 11,contains at least six important resistance genes,but these genes have not been widely employed in resistance breeding since existing markers hardly satisfy current breeding needs due to their limited scope of application.In this study,two PCR-based markers,Pikp-Del and Pi1-In,were developed to target the specific In Del(insertion/deletion)of the Pik-p and Pi-1 genes,respectively.The two markers precisely distinguished Pik-p,Pi-1,and the K-type alleles at the Pik locus,which is a necessary element for functional genes from rice varieties.Results also revealed that only several old varieties contain the two genes,of which nearly half carry the K-type alleles.Therefore,these identified varieties can serve as new gene sources for developing blast resistant rice.The two newly developed markers will be highly useful for the use of Pik-p,Pi-1 and other resistance genes at the Pik locus in markerassisted selection(MAS)breeding programs.

Synthesis,Crystal Structure and Insecticidal Activity of N-(pyridin-2-ylmethyl)-1-phenyl-1,4,5,6,7,8-hexahydrocyclohepta[c]pyrazole-3-carboxamide

The title compound N-（pyridin-2-ylmethyl）- 1-phenyl- 1,4,5,6,7,8-hexahydrocy-clohepta[c]pyrazole-3-carboxamide 5 （C21I-I22N40, Mr = 346.42） has been synthesized andstructurally characterized by IR, 1H NMR, 13C NMR, H RMS and single-crystal X-raydiffraction. The crystal crystallizes in monoclinic system, space group P21/n with a = 8.668（2）,b = 22.236（4）, c = 9.539（2） A, β = 108.68（3）°, V = 1786.4（7）/k3, Z = 4, Dx= 1.288 g/cm3,F（000） = 736,μ（MoKa） = 0.649 mm^-1, the final R = 0.0354 and wR = 0.0933 with 3234observed reflections with I 〉 2σ（/）. The benzene and pyrazole rings are nearly coplanar with adihedral angle of 50.977（46）°. The dihedral angle between the central pyrazole and pyridinerings is 11.688（46）°. No classical hydrogen bonds were found in the molecules. Two adjacentmolecules in crystal packing of compound 5 were linked by two intramolecularhydrogen-bonding interactions C（15）-H（15）…O（1） to generate a stable structure. Compound 5had weak insecticidal activity against the diamondback moth （Plutella xylostella）, butexhibited good activity against cotton bollworm （Helicoverpa armigera）.

Rice sl-MH-1 mutant induces cell death and confers blast resistance via the synergistic roles of signaling systems

Serotonin is ubiquitous across all forms of life and functions in responses to biotic and abiotic stresses.In rice,the conversion of tryptamine to serotonin is catalyzed by Sekiguchi lesion(SL).Previous studies have identified an sl mutation(a null mutation of SL)in several rice varieties and confirmed its increase of resistance and cell death.However,a systematic understanding of the reprogrammed cellular processes causing cell death and resistance is lacking.We performed a multi-omics analysis to clarify the fundamental mechanisms at the protein,gene transcript,and metabolite levels.We found that cell death and Magnaporthe oryzae(M.oryzae)infection of the sl-MH-1 mutant activated plant hormone signal transduction involving salicylic acid(SA),jasmonic acid(JA),and abscisic acid(ABA)in multiple regulatory layers.We characterized the dynamic changes of several key hormone levels during disease progression and under the cell death conditions and showed that SA and JA positively regulated rice cell death and disease resistance.SL-overexpressing lines confirmed that the sl-MH-1 mutant positively regulated rice resistance to M.oryzae.Our studies shed light on cell death and facilitate further mechanistic dissection of programmed cell death in rice.

Optimization of micropropagation and <i>Agrobacterium</i>-mediated gene transformation to spinach (<i>Spinacia oleracea</i>L.)

Spinach is one of the dioecious plant which is considered as a model plant in genetic and molecular studies of sex determination because of its special characteristics such as low chromosome number and short life cycle. An efficient protocol for Spinacia oleracea Agrobacterium-mediated gene transformation was developed. The leaf disks, roots, hypocotyls and cotyledons of this plant were inoculated with LBA4404. LBA4404 carrying pCAMBIA3301 binary vector with 35SCaMV gusint and 35SCaMV bar cassettes. Effects of two preparation condition (induction of vir genes and noninduction) were considered. Also effects of different number days of co-cultivation and pre-culture of explants were examined. After co-cultivation, the explants were transferred to regeneration medium containing 250 mg·L-1 Carbeniciline. Transient expression efficiency was calculated based on the number of blue spots per explants one week after inoculation. Based on the results of transient expression, stable transformation was carried out. After formation of callus the histochemical GUS assay was carried out on some parts of them and other parts were leaved for being regenerated.

Distinguished discriminatory separation of CO2 from its methane-containing gas mixture via PEBAX mixed matrix membrane

Highly selective separation of CO_2 from its methane-containing binary gas mixture can be achieved by using Poly(ether-block-amide)(PEBAX)mixed matrix membranes(MMMs).According to FESEM and AFM analyses,silica-based nanoparticles were homogenously integrated within the polymer matrix,facilitating penetration of CO_2 through the membrane while acting as barrier for methane gas.The membrane containing 4.6 wt% fumed silica(FS)(PEBAX/4.6 wt%FS)exhibits astonishing selectivity results where binary gas mixture of CO_2/CH_4 was used as feed gas.As detected by gas chromatography,in the permeate side,data showed a significant increase of CO_2 permeance,while CH_4 transport through the mixed matrix membrane was not detectable.Moreover,PEBAX/4.6 wt%FS greatly exceeds the Robeson limit.According to data reported on CO_2/CH_4 gas pair separation in the literature,the results achieved in this work are beyond those data reported in the literature,particularly when PEBAX/4.6 wt%FS membrane was utilized.

Single-cell RNA sequencing of mitoticarrested prospermatogonia with DAZL::GFP chickens and revealing unique epigenetic reprogramming of chickens

Background:Germ cell mitotic arrest is conserved in many vertebrates,including birds,although the time of entry or exit into quiescence phase differs.Mitotic arrest is essential for the normal differentiation of male germ cells into spermatogonia and accompanies epigenetic reprogramming and meiosis inhibition from embryonic development to post-hatch.However,mitotic arrest was not well studied in chickens because of the difficulty in obtaining pure germ cells from relevant developmental stage.Results:We performed single-cell RNA sequencing to investigate transcriptional dynamics of male germ cells during mitotic arrest in DAZL::GFP chickens.Using differentially expressed gene analysis and K-means clustering to analyze cells at different developmental stages(E12,E16,and hatch),we found that metabolic and signaling pathways were regulated,and that the epigenome was reprogrammed during mitotic arrest.In particular,we found that histone H3K9 and H3K14 acetylation(by HDAC2)and DNA demethylation(by DNMT3B and HELLS)led to a transcriptionally permissive chromatin state.Furthermore,we found that global DNA demethylation occurred gradually after the onset of mitotic arrest,indicating that the epigenetic-reprogramming schedule of the chicken genome differs from that of the mammalian genome.DNA hypomethylation persisted after hatching,and methylation was slowly re-established 3 weeks later.Conclusions:We found a unique epigenetic-reprogramming schedule of mitotic-arrested chicken prospermatogonia and prolonged hypomethylation after hatching.This will provide a foundation for understanding the process of germ-cell epigenetic regulation in several species for which this process is not clearly described.Our findings on the biological processes related to sex-specific differentiation of prospermatogonia could help studying germline development in vitro more elaborately.

Identification of Genetic Relationship of Hybrid Rice Varieties to Their Parents with PCR Products

[Objectives]To establish a simple,rapid and accurate method for identifying the genetic relationship of hybrid rice varieties to their parents.[Methods]Taking F1 hybrids Liangyou 336,Deliangyou Huazhan,and the parents of Liangyou 336,i.e.,C815S(♀)and R336(♂),as experimental materials,the genetic relationship of the hybrid rice varieties to the parental materials was identified by way of PCR amplification with the 48 pairs of SSR primers of Protocol for Identification of Rice Varieties:SSR Marker Method(NY/T 1433-2014).[Results]The genetic relationship of the hybrid rice varieties could be determined by comparing the PCR amplification products of the mixed DNA of the parents and the DNA of the F1 hybrids.[Conclusions]This method not only reduced the number of samples required but also had a good visual effect and high accuracy.