Mapping of Purple Gene in Spears of Asparagus(Asparagus officinalis L.)

[Objectives]This study was conducted to find out regulatory genes related to purple in spears of asparagus(Asparagus officinalis L.).[Methods]The stable asparagus inbred line JX1513-5(the base of the spear is purple)and JLV1718-7(the base of the spear is green)were used as parents to study the genetic law of purple/green traits in their offspring.[Results]The results showed that the purple in the basal part of asparagus spear was controlled by a pair of alleles,and purple was dominant over green.The F 2 segregation population was resequenced by the bulk segregation analysis(BSA)method,and the purple trait in the basal part of asparagus spear was located in the interval of 24.51-25.08 Mb on Chr07 chromosome,which included 47 genes.According to the annotation information,three candidate genes were screened out:LOC109849403,LOC109849430 and LOC109849442.The candidate genes were verified by real-time fluorescence quantitative PCR(qRT-PCR),and finally LOC109849442 was obtained as the candidate gene for controlling the purple/green trait in the basal part of asparagus spear.[Conclusions]This study lays a foundation for the breeding of new asparagus varieties and molecular marker-assisted breeding.

Technical Efficiency of"Master Switch"in Widening the Human Living Space by Fenlong Tillage

This paper reports"the fourth set"of drilling tillage after"ploughing(hoeing)"by manpower,animal power and tractor——Fenlong tillage technology(referred to as"Fenlong technology").It makes the cultivated land and saline alkali land transformed into"sponge"farmland through"drill bit"vertically digging into the soil,one-time deep cultivation,not disturbing soil layer,and granulating the soil,to bring about the effects of"expanding the land,releasing the natural force,water conservation,disaster reduction,carbon reduction,tillage saving,fertilizer saving and grain increasing",and can broaden the space for human survival and development.It has been applied to more than 50 crops in 28 provinces,including Guangxi,Hebei and Tibet.Without increasing the use of fertilizer and water,the yield of varieties in cultivated land increased by 10%-50%,while that in saline and alkaline land increased by 20%-100%.The storage of natural precipitation increased by 100%,and drought,high temperature and low temperature decreased by 20%-30%,and carbon sequestration and emission reduction reached 20%.It is proposed that China can implement the"dual strategy"platform of farmland,saline alkali land,rivers and water bodies constructing the"incremental""five new warehouses"of grain,water,fish,sugar,etc.and"increased grain return"abroad,to ensure national security.

Polyploidy events shaped the expansion of transcription factors in Cucurbitaceae and exploitation of genes for tendril development

Cucurbitaceae is one of the most important plant families distributed worldwide.Transcription factors(TFs)regulate plant growth at the transcription level.Here,we performed a systematic analysis of 42641 TFs from 63 families in 14 Cucurbitaceae and 10 non-cucurbit species.Whole-genome duplication(WGD)was the dominant event type in almost all Cucurbitaceae plants.The TF families were divided into 1210 orthogroups(OGs),of which,112 were unique to Cucurbitaceae.Although the loss of several gene families was detected in Cucurbitaceae,the gene families expanded in five species that experienced a WGD event comparing with grape.Our findings revealed that the recent WGD events that had occurred in Cucurbitaceae played important roles in the expansion of most TF families.The functional enrichment analysis of the genes that significantly expanded or contracted uncovered five gene families,AUX/IAA,NAC,NBS,HB,and NF-YB.Finally,we conducted a comprehensive analysis of the TCP gene family and identified 16 tendril-related(TEN)genes in 11 Cucurbitaceae species.Interestingly,the characteristic sequence changed from CNNFYFP to CNNFYLP in the TEN gene(Bhi06M000087)of Benincasa hispida.Furthermore,we identified a new characteristic sequence,YNN,which could be used for TEN gene exploitation in Cucurbitaceae.In conclusion,this study will serve as a reference for studying the relationship between gene family evolution and genome duplication.Moreover,it will provide rich genetic resources for functional Cucurbitaceae studies in the future.

Gene characterization and molecular pathway analysis of reverse thermosensitive genic male sterility in eggplant(Solanum melongena L.)

The naturally occurring mutant eggplant line 05ms was identified with reverse thermosensitive genic male sterility(rTGMS),but its temperature-responsive fertility mechanisms remain largely unknown.Here,we studied the flower morphology,anther cellular structure,and genome-wide gene expression of this rTGMS line.Candidate genes for thermosensitive male sterility during the microspore development of 05ms and the temperature-insensitive line S63 under low-temperature(LT)and high-temperature(HT)conditions were identified.Under LT,tapetum cells were vacuolated and had delayed disintegration in 05ms.RNA-seq analysis indicated that DEGs were enriched in the KEGG pathways‘plant hormone signal transduction’,‘starch and sucrose metabolism’,and‘phenylpropanoid biosynthesis’.We identified two genes,4CLL1(Sme2.5_00368.1_g00010.1)and CKI1(Sme2.5_10056.1_g00002.1),which could potentially regulate eggplant anther development and may be candidate genes for rTGMS.Finally,we propose a working model of anther abortion for rTGMS in eggplant.CKI1 responds to LT stress and causes expression changes in genes related to anther development,such as 4CLL1,and the cellular structure of the tapetum becomes abnormal,causing male sterility.The findings of this study explain the underlying molecular mechanisms of male sterility in eggplant rTGMS lines.

Characterization of Gh SERK2 and its expression associated with somatic embryogenesis and hormones level in Upland cotton

Somatic embryogenesis （SE） is one of the most important steps during regeneration of cotton, but the molecular mechanism of SE remains unclear. SOMATIC EMBRYOGENSIS RECEPTOR KINASE （SERK） gene is known to function in SE. A homolog GhSERK2 （accession number： JF430801） was cloned from Upland cotton and characterized for its functions in SE. GhSERK2 expressed in different tissues and showed higher expression level in floral organs than vegetative ones with the highest levels in ovule and anther. GhSERK2 expressed during SE with a high level at globular embryos stage. Upon treatment with indole-3-butytic acid （IBA）, the transcription level of GhSERK2 was induced and promoted SE subsequently. A 2-day treatment of 2,4-dichlorophenoxyacetic acid （2,4-D） induced the expression of GhSERK2, but treatments of 2,4-D for longer periods sharply inhibited the GhSERK2 transcription level of embryogenic callus （EC）. The levels of hormones, including 3-indoleacetic acid （IAA）, abscisic acid （ABA）, and brassinosteroid （BR）, were increased in the initial calli induced from the over-expression of GhSERK2 cotton. Our results indicated that GhSERK2 expression was associated with induction of SE and closely related to hormone levels during tissue culture in Upland cotton, and the gene might play an important role in regeneration of cotton.

Research of Protected Cultivation Techniques of Toona sinensis on Mountain Slopes

In order to prolong the supply period and increase the economic value of Toona sinensis,the protected cultivation of T. sinensis was carried out on mountainous slope to achieve stagger-season planting and the year round supply. The results showed that in late October to midNovember,the temperature was moderate( 1-10℃) and suitable for the lifting of seedlings and temporary planting dormancy. The line spacing of rooted seedlings in greenhouse was( 22-24) cm ×( 4-5) cm and planting density was 900 000-1 200 000 seedlings/ha. The line spacing of seedlings was( 14-13) cm ×( 3-4) cm and planting density was 1 950 000-2 100 000 seedlings/ha. T. sinensis began to sprout at 8-10℃ and stretch at 10-12℃. When the temperature was stable at 14℃ or above,T. sinensis started bolting and emerging leaves. The terminal buds sprouted in 40-50 d for the T. sinensis planted in greenhouse with the day temperature of 15-22℃ and night temperature over 10℃; the terminal buds sprouted in about 15 d for those planted in the greenhouse with day temperature of 20-25℃ and night temperature over 10℃. T. sinensis could be picked 3 times when planted in greenhouse,and the yield was about 24 720 kg/ha.

Breeding of a New Clubroot-resistant Chinese Cabbage Variety, CCR11242

In this study, using Japanese radish cytoplasmic sterile line ＂Half Green Cabbage＂ （HGC） and clubfoot-resistant South Korean cabbage variety ＂ CR Cabbage King＂ （CCK） as experimental materials, serf-incompatible DH lines were obtained by mierosporo culture. Through five gnerations of backeross, CCR11239, a clubroot-resistant radish cytoplasmic sterile line, was obtained. Through six generations of serf-crossing, CCR11240, the maintainer line of CCR11239, was bred. After the cross of CCK and Luchunbai 1 （83 - 1 ） and five generations of self-crossing, serf-compatible line CCRl1241 was obtained. Through crossing CCR11240 with CCRl1241, the new cabbage variety CCRl1242 was obtained. After variety comparison test, provincial regional test and production demonstration test, the new variety was registered in 2012. The average yield of CCR11242 reached 60 975 kg/hm2, which was improved by 142% compared with Luchunbai 1 （83 - 1 ）. The disease index of CCR11242 was 5.63, which was 88.13 lower than the control, indicating high resistance （HR） to clubfoot.

Analysis of short fruiting branch gene and Marker-assisted selection with SNP linked to its trait in upland cotton

Background： With the rapid development of genomics, many functional genes have been targeted. Molecular marker assisted selection can accelerate the breeding process by linking selection to functional genes. Methods： In a study of upland cotton （Gossypium hirsutum L.）, the F2 segregated population was constructed by crossing X1570 （short branches） with Ekangmian 13 （long branches） to identify the short fruiting branch gene and marker assisted selection with SNP（Single Nucleotide Polymorphisms, SNP） linked to its trait. Result： The result demonstrated that linked SSR marker BNL3232 was screened by BSA（Bulked segregant analysis, BSA） method; one SNP locus was found, which was totally separated from the fruiting branches trait in upland cotton. Conclusion： It was verified that this SNP marker could be used for molecular assisted selection of cotton architecture

Identification of Nucleus Sterility Candidate Genes Using a Resequencing Technique in Sweet Pepper Sterile Line AB91

Breeding hybrids with nuclear malesterile lines is an important method for the cross-breeding of sweet peppers. To date, few reports have been published on the nuclear malesterility gene of sweet pepper. Yet, there are approximately 20 pepper nuclear malesterility lines in the world. Using the self-developed testing material, sweet pepper nuclear malesterile dual-purpose line AB91, the genome-wide resequencing technique was applied to find that the mutation site causing the abortion of sweet pepper nuclear malesterility AB91 is on chromosome #5. The mutation gene Capana05g000747 was filtered out and validated by the flight mass spectrometry genotyping and quantitative realtime PCR method and determined to be the gene causing the abortion of sweet pepper nuclear male sterility AB91. The gene Capana05g000747 mutation site is a non-synonymous mutation site located at the 6th exon, the base C mutated into A, and the amino acid changed from alanine to serine. The three-dimensional protein structure of fertile and sterile plant Capana05g000747 was predicted. The results showed that the three-dimensional structure of the two proteins differed significantly. Sequence alignment analysis showed that the gene Capana05g000747 has a similar function to gene At2g02148. The gene At2g02148 contains a pentatricopeptide repeat protein which has important physiological functions in the gene expression process of organelles and is closely related to the performance of malesterility genes. Therefore, Capana05g000747 was selected as an important candidate gene for sweet pepper nuclear male sterile testing material AB91.

Metabolomics Analysis of Metabolites in Forsythia suspense Fruit Using UPLC/ESI-Q TRAP-MS/MS

Forsythiae Fructus,the fruit of Forsythia suspense is a traditional Chinese hebal medicine that has the antiviral and antioxidant effects in China.Modern analytical chemistry studies showed that the extracts of Forsythiae Fructus contain many bioactive components,such as flavonoids,lignans,phenolic acids,and terpenoids,which can be used to anti-inflammatory and treat toxicity,tonsillitis,ulcers,pharyngitis and acute nephritis.In order to study the types and quantities of metabolites in Forsythiae Fructus,we isolated,identified and analysed metabolites between two varieties of Forsythiae Fructus using UPLC/ESI-Q TRAP-MS/MS.The results showed that a total of 407 metabolites were identified in Forsythiae Fructus using UPLC/ESI-Q TRAP-MS/MS,including 21 terpenoids,68 phenolic acids,63 flavonoids,43 amino acids and derivatives,22 alkaloids,55 lipids,24 lignans and coumarins,31 nucleotides and derivatives,29 organic acids,and 51 other metabolites.Among,lignans and coumarins,terpenoids,organic acids,lipids,and phenolic acids were rich in Forsythiae Fructus,which accounted for more than 60%of the total metabolite content.Differential metabolite analysis revealed that 80 metabolites differed significantly between the two types of Forsythiae Fructus.Our results greatly enrich the Forsythiae Fructus phytochemical composition database and provide valuable information for further study of the metabolites of Forsythiae Fructus.

Hardness Genetic Analysis of Pink Hard-fruit Tomato

Hardness genetic laws of pink hard-fruit tomato were studied for breeding new varieties of pink hard-fruit tomato. The change of hardness degree and heredi- tary characters of tomato were studied using the texture analyzer, with female parents P27, male parents P31 and tomato F1 combination 15# as the test materials. The result showed that transverse hardness was the lowest, followed by longitudinal hardness, and the hardness of fruit shoulder was the highest. The average hardness at green fruit stage was higher, the hardness at turning-color period and red ripe stage decreased significantly. The average hardness of P27, P31, F1 combination 15# at red ripe fruit was 13.27, 10.88, 12.28 N, respectively. The hardness at red fruit stage and green fruit stage had significantly positive correlation. Fruit shape index of F1 combinations 15#at red ripe stage was 0.87. The fruit shape was round with 5-6 locules. The pericarp thickness was 0.7-0.8 cm. The single fruit weight was 258 g. It was the ideal bred variety for pink hard-tomato fruit. There was no necessary connection among fruit hardness and fruit size, pericarp thickness, ventricle numbers, single fruit weight. Fruit shape index could be used as indirect selection index of the hardness of tomato.

Discussions of Exploring"41635"Potential of Food for More than 400 Million People through Developing"Fenlong Agriculture"

This paper discussed exploring"41635"potential of food for more than 400 million people through developing"Fenlong Agriculture"in China.It clarified that"5.0 Fenlong Technology"is composed of"drill type Fenlong tillage tools+Fenlong farming technology+Fenlong crop cultivation technology standards",and has coupling and interaction"common key technologies in agricultural production,resource utilization and ecological improvement,and"Great Science"connotation and characteristics,the yield increase rate for 40 kinds of crops in 26 provinces in China reached 10%-150%.China uses"5.0"Fenlong Technology"to develop"Fenlong agriculture"to implement the optimization and transformation of 67 million ha of cultivated land,13 million ha of saline-alkali land,and 67 million ha of degraded grassland and related farming systems,excavating its new round of multiple activations and increasing the huge concentration of"gifted resources"such as soil,water,gas,temperature and light,develop"five systems","big pattern","rainfed and environment-friendly"and other types of Fenlong agriculture,so as to increase food,meat,fish and other food sources to feed 400 million people every year,increase natural soil storage by 100 billion m3,reduce groundwater collection by 60 billion m3,reduce the use of chemical fertilizers and pesticides by more than 30%,the comprehensive benefits including yield increase,quality improvement,water saving,chemical fertilizer reduction,ecological protection,and disaster reduction can increase by more than 50%(these are 41635 potential),to realize the objective that a place can feed its people by its own resources.

Indoor Inoculation and Identification Technology of Sweet (Hot) Pepper Virus

[Objective] The paper was to establish an affordable indoor virus inoculation and identification technology. [Method] Taking Tobacco Mosaic Virus （TMV） and Cucumber Mosaic Vires （CMV） of sweet （hot） pepper as the sources of virus, an affordable indoor virus inoculation and identification technology was developed in the paper. [ Result] The suitable inoculation concentration of CMV was five to ten times, and the best seedling age for inoculation was five to six leav- es. Suitable inoculation concentration of TMV was 20 to 30 times, and the best seedling age for inoculation was three to six leaves. Single inoculation technology was mainly used for indoor virus inoculation and identification of sweet （hot） pepper, and complex inoculation technology could also be adopted with first, st inoculation of CMV and late inoculation of TMV. For mixed inoculation technology, CMV： TMV should be 1： 1. Complex inoculation and mix inoculation should base on the tech- nology of single inoculation. Disease resistant materials, AID1-W22-dg176, ABgl-W22-48123, AB91-DL-6428, HY031-2-8-1-6, BYT-4-1-3-6-8, JFG-2-1-2-6, JF8S-1-1-5-4-8 and I＇502-1-1-3-5, were identified by this method. [ Conclusion] This research provided scientific basis for standardization of indoor inoculation of sweet （hot） pepper virus.

Polygenic Heritability of Rose Root Rot Disease Resistance in Offspring of Rosa multiflora

Rose root rot disease caused by Pythium helicoides Drechsler has brought heavy damage to cut rose production in Japan,and it has became apparent that Rosa multiflora showed resistance to this disease.Roses have been bred by interspecies crossing with Rosa species including of R.multiflora,and many progenies of R.multiflora have been bred as determined cluster-flowered roses.However,the genetic architecture of this resistance remains.So,in this study,four kinds of R.multiflora and 21 rose cultivars that have been clarified in the cross-fertilization genealogy with R.multiflora were used as plant materials,and the inheritability of resistance to root rot disease was investigated.Four kinds of R.multiflora had all high resistance,and the resistance to root rot disease in R.multiflora was inherent characteristic of species.Five varieties not less than 50% in relatedness with R.multiflora were shown as varied ‘resistant’ and ‘moderately resistant’.Therefore,the inherent resistant characteristic in R.multiflora was not controlled by single major gene,and the resistibility in R.multiflora was not cytoplasmic but nuclear heredity.Although there was significant negative relation between relative root rot severity and relatedness with R.multiflora,the variance of relative root rot severity had tendency to expand as relatedness with R.multiflora reduced.This result,therefore,indicated that the contingency of resistance expression increased as the relatedness with R.multiflora was reduced.The varieties having low degree of relatedness with R.multiflora had similar relative root rot severity with R.multiflora.These results indicated that we would be able to select resistant roses from progenies crossed with R.multiflora.

Status of Agricultural Production and Crop Variety Improvement in Thailand

We introduced basic conditions of agricultural production in Thailand, and variety improvement of major crops, including rice, cassava, rubber, and vegetable, in the hope of providing reference for agricultural production and crop variety improvement in Hubei Province and even in the whole country.

Study on Available Nutrients of Soil in Fenlong by a New Farming Method

Taking 0-30 cm, 0-40 cm, 0-50 cm, 0-60 cm of dry land Fenlong and 0-40 cm paddy field as samples of soil profile with corresponding original soil as the control group, we measured content of organic matter, quick-acting N, quick-acting P and quick-acting K, effective B, Cu, Zn and Mn. Results indicate that available nutrients in soil after Fenlong were higher than original soil. For dry land, the increase of organic matter, quick-acting N, quick-acting P and quick-acting K is 3.02%-35.16%, 6.80%-39.54%, 2.81%-44.46%, and 7.72%-53.71% respectively. There is also increase in effective content of trace element, B, Cu, Zn and Mn. For paddy field, the increase of organic matter, quick-acting N, P and K is 19.64%, 24.02%, 24.27% and 57.78% respectively. Besides, there is also increase in content of trace element, B, Cu, and Zn. On the basis of analysis, we put forward the new theory of crop cultivation "root" theory.

Analysis of the Current Structure and Response Strategies of Intensive Vegetable Seedling Cultivation in Hebei Province

In the last few years, vegetable industry developed rapidly in Hebei province, intensive raising seedling as its main component, also entered a period of rapid development. The background and characteristics, the current situation and existed questions of intensive seedling cultivation were analyzed. A series of developmental ideas and response strategies for developing more quickly in Hebei Province were put forward.

Study on Compositions of Grain Starch and SGP-1 Protein in Black Grain Wheat

In view of the lack of wheat genetic resources with high amylose and high resistant starch contents in the present world,the grain starch components and SGP-1 (SGP-A1,SGP-B1 and SGP-D1) protein composition of 43 black-kernel wheat genetic resources such as Jizi 439 were identified by SDS-PAGE electrophoresis detection.The results showed that 11 materials were lack of SGP-A1,and no materials had SGP-B1 and SGP-D1 deletion.Seven materials were identified to have an amylose content more than 30% of the total starch.A total of 12 materials were selected for the determination of resistant starch content,and five materials were found to have a high resistant starch content.The above results lay a foundation for wheat breeding for high resistant starch content.

Physiological Response and Transcriptome Analysis of Cotton Leaves under Low Temperature Stress at the Two-leaf Stage

[Objectives]To investigate the effect of low temperature treatment on cotton leaves at the two-leaf stage.[Methods]Two cotton varieties(CN01 and SJB016(low temperature-tolerant))were used as the trial materials.They were treated at 25(CK)and 12℃(low temperature)for 0,3,6,12,24,48 and 72 h,respectively.Then,the changes in the contents of MDA,SS and Pro in the cotton leaves were analyzed.Based on the analysis results,RNA-seq verification was performed.[Results]Two cotton varieties(CN01 and SJB016(low temperature-tolerant))were used as the trial materials.They were treated at 25(CK)and 12℃(low temperature)for 0,3,6,12,24,48 and 72 h,respectively.Then,the changes in the contents of MDA,SS and Pro in the cotton leaves were analyzed.Based on the analysis results,RNA-seq verification was performed.[Conclusions]These genes may play an important role in improving the cold resistance of cotton.

Effects of Salt Stress on Growth and Ion Distribution in Seedlings of Asparagus officinalis L.

An Asparagus officinalis L. cultivar NJ978 was used to study the growing rate, salt ion content and mineral ion uptake and distribution in plants under salt stress. The results showed that salt stress significantly inhibited seedling growth and there was a negative relationship between seedling growth and salt concentration. The seedlings growth showed no significant decrease under low salt stress （NaCl≤50 mmol/L）, while high concentrations of NaCl led to detrimental effects on the growth of A. officinalis L.. With the increase of NaCl concentration, the Na ＋ content increased gradually in A. officinalis L. roots, stems and leaves while the K^＋ and Ca^2＋ contents had slow decreases. Under low salt stress, the seedlings of A. officinalis L. could prevent Na ＋ from transporting to aerial part by withholding Na ＋ in root, which could keep the balance of ions in aerial part, but under high salt stress （NaCl 〉 100 mmol/L）, the aerial part of seedlings accumulated superabundance of Na ＋, which limited the uptake of K^＋ and Ca^2＋ , so seedlings were damaged heavily, which manifested as the gradual decrease of K^＋/Na ＋ and Ca^2＋ /Na ＋ in the roots and aerial part of the seedlings. The selective absorption of K^＋, Ca^2＋ and Na ＋ （AS K,Na and AS Ca,Na ） was significantly increased with increased salinity, while the selective transportation （TS K,Na and TS Ca,Na ） increased at first and then decreased with the increased salinity. Therefore, the strong ability of salt exclusion and the regionalized distribution of ions in roots, stems and leaves could be one of the salt tolerance mechanisms of A. officinalis L.