Selection of Homozygous Cotton Lines Transformed with Two Insect-Resistant Genes

A plant expression vector containing a chimeric Bt29K gene coding for the activated Cry1Ac protein and the arrowhead proteinase inhibitior gene API B were introduced into the cotton cultivar Jihe321 mediated by Agrobactertium tumefaciens. Based on the results of kanamycin resistant testing, PCR detection for both foreign genes and insect bioassay using Heliethis armigera , nine transgenic homozygous cotton lines with insect resistance of more than 90% and better agronomic traits were bred through six generations from the original transgenic plants. Results from insect bioassay and sequence analysis of the PCR products of plants from some homozygous lines indicated that the chimeric Bt29K gene was stably inherited in these transgenic cotton lines. The main agronomic characters of these homozygous cotton lines, such as boll productivity and fibre strength, were better than that of the original cotton cv. Jihe321.

Expression analysis of the R2R3-MYB gene family in upland cotton and functional study of GhMYB3D5 in regulating Verticillium wilt resistance

Improving plant resistance to Verticillium wilt(VW),which causes massive losses in Gossypium hirsutum,is a global challenge.Crop plants need to efficiently allocate their limited energy resources to maintain a balance between growth and defense.However,few transcriptional regulators specifically respond to Verticillium dahliae and the underlying mechanism has not been identified in cotton.In this study,we found that the that expression of most R2R3-MYB members in cotton is significantly changed by V.dahliae infection relative to the other MYB types.One novel R2R3-MYB transcription factor(TF)that specifically responds to V.dahliae,GhMYB3D5,was identified.GhMYB3D5 was not expressed in 15 cotton tissues under normal conditions,but it was dramatically induced by V.dahliae stress.We functionally characterized its positive role and underlying mechanism in VW resistance.Upon V.dahliae infection,the up-regulated GhMYB3D5 bound to the GhADH1 promoter and activated GhADH1expression.In addition,GhMYB3D5 physically interacted with GhADH1 and further enhanced the transcriptional activation of GhADH1.Consequently,the transcriptional regulatory module GhMYB3D5-GhADH1 then promoted lignin accumulation by improving the transcriptional levels of genes related to lignin biosynthesis(GhPAL,GhC4H,Gh4CL,and GhPOD/GhLAC)in cotton,thereby enhancing cotton VW resistance.Our results demonstrated that the GhMYB3D5 promotes defense-induced lignin accumulation,which can be regarded as an effective way to orchestrate plant immunity and growth.

Development and identification of Verticillium wilt-resistant upland cotton accessions by pyramiding QTL related to resistance

Cotton Verticillium wilt is a serious soil-borne disease that leads to significant losses in fiber yield and quality worldwide. Currently, the most effective way to increase Verticillium wilt resistance is to develop new resistant cotton varieties. Lines 5026 and 60182 are two Verticillium wilt-resistant upland cotton accessions. We previously identified a total of 25 quantitative trait loci（QTLs） related to Verticillium wilt resistance from 5026 and 60182 by assembling segregating populations from hybridization with susceptible parents. In the current study, using 13 microsatellite markers flanking QTLs related to Verticillium wilt resistance, we developed 155 cotton inbred lines by pyramiding different QTLs related to Verticillium wilt resistance from a filial generation produced by crossing 5026 and 60182. By examining each allele＇s effect and performing multiple comparison analysis, we detected four elite QTLs/alleles（q-5/NAU905-2, q-6/NAU2754-2, q-8/NAU3053-1 and q-13/NAU6598-1） significant for Verticillium wilt resistance, pyramiding these elite alleles increased the disease resistance of inbred lines. Furthermore, we selected 34 elite inbred lines, including five lines simultaneously performing elite fiber quality, high yield and resistance to V. dahliae, 14 lines with elite fiber quality and disease resistance, three lines with high yield and disease resistance, and 12 lines with resistance to V. dahliae. No correlation between Verticillium wilt resistance and fiber quality traits/yield and its components was detected in the 155 developed inbred lines. Our results provide candidate markers for disease resistance for use in marker-assisted breeding（MAS）, as well as elite germplasms for improving important agronomic traits via modern cotton breeding.

Wrinkle Resistant Finishing of Cotton Fabrics with the Complex System of PBTCA/CA

In this paper,the wrinkle resistant finishing of cotton fabric with the complex system of PBTCA and CA was mainly discussed.The influence of finishing conditions such as the amount of finishing agent and catalyst,curing temperature and curing time were studied.Wrinkle recovery angle(WRA),breaking strength(BS) and whiteness(Wh) of finished fabric were measured.The experimental data indicated that the best finishing conditions were PBTCA 120g/L,CA 50g/L,sodium hypophosphite 50g/L,triethanolamine 20g/L,cured 170℃ for 120s.

Combining Ability and Heterosis Between High Strength Lines and Transgenic Bt (Bacillus thuringiensis) Bollworm-Resistant Lines in Upland Cotton (Gossypium hirsutum L.)

To analyse the combining ability and heterosis between high-strength lines and transgenic Bt bollworm-resistant lines in upland cotton, 5 high-strength lines were crossed as female lines with 12 transgenic Bt bollworm-resistant lines according to NCII design. It was demonstrated that the compositions of variance in various traits were quite different. For seed cotton yield, lint yield, boll numbers per plant and boll weight, the dominant (special combining ability) effects were the major effects, accounting for 87.38, 84. 40, 80. 04 and 64. 46% of the total phenotypic variances, respectively, while for fibre strength and micronaire value, the additive (general combining ability) effects had the major effects, with a ratio of additive variance to phenotypic variance of 78.85 and 43.80%. As for lint percent and 2.5% span length, the dominant and additive variances had similar effects, in phenotypic variances (54.94 and 40.11% for lint percent, 45.76 and 42.49% for 2. 5% span length, respectively). The mid-parent heterosis (Hpm), surpassing parent heterosis (Hpb) and competitive heterosis (Hck) for seed cotton yield and lint yield were both extremely significant. For fibre properties, the Hck and Hpm of 2.5 % fibre span length were extremely significant, the Hck of fibre strength was significant, and the favorable negative Hck of micronaire was also extremely significant. The increments of hybrid over common variety were 17% for lint yield and fibre strength, 7% for fibre span length, and 4 % for fineness.

Cloning of Partial Sodium Channel Gene From Strains of Fenvalerate-Resistant and Susceptible Cotton Aphid(Aphis gossypii Glover)

The strain of fenvalerate-resistant cotton aphids was selected using fenvalerate insecticide in the laboratory, the resistance factor of the strain was 199.54. Three degenerate primers were designed and used to perform PCR amplification. A cDNA encoding partial sodium channel gene was cloned from the fenvalerate-resistant and -susceptible strains. There were two nucleotide acid differences between fenvalerate-resistant strain and -susceptible strain, resulting in an amino acid mutation, Met→Leu. It is predicted that the mutation is related to the cotton aphid resistance to fenvalerate.

Breeding of High Yield,High Quality and Disease-Resistant Hybrid Cotton Varieties Hanza 160 and Hanza 1692

The hybrid cotton varieties of Hanza 160 and Hanza 1692 were bred by reciprocal crosses between Han 5158 and Han 333 in Hainan and Hebei for several years. Both varieties had the characters of high yield, high quality and disease resistance. Hanza 160 and Hanza 1692 were approved by Hebei Crop Variety Approval Committee in 2013 and 2016, respectively.

Breeding of Good-quality High-yield Salt-tolerant and Drought-resistant Hybrid Cotton Variety Hengyou 12

Salt and drought tolerance, high yield and quality, disease and pest resistance of cotton varieties are new requirements for the current situation in cotton production. Hengyou 12 is a transgenic Bt cotton variety bred by hybridizing JD 28 （interspeciflc progeny of upland cotton, sea-island cotton, wild cotton species） as female parent with GK55 （transgenic Bt cotton variety） as male parent. The combining of parents was based on the complementary principle of parental traits. Hengyou 12 with strong heterosis was selected under disease stress, pest stress, salt stress and drought stress. This variety has many excellent characteristics, such as good resis- tance to diseases and pests, high yield, excellent quality, strong drought resistance and salt tolerance. ＇Hengyou12＇ was approved by Hebei Province in 2015, and is suitable for planting in middle and south areas of Hebei and similar ecological areas in the Yellow River basin.

Studies on Cotton Breeding Resistant to Fusarium and Verticillium wilt Diseases

Both Fusarium and Verticillium wilts are important soil-borne diseases,which can not be effectively controlled by chemical fungicides.The two diseases,especially Verticillium wilt,have

Insect Resistance of Different Tissues of Transgenic Cotton to Spodoptera exigua(Hbner)

[Objective] The aim was to learn the resistance of different tissues and organs of transgenic cotton to Spodoptera exigua （Hbner）. [Method] Flowers,the 1st,the 3rd,the 6th and the 14th leaves from the top of 33B,GK12 and SGK321 were used to feed S. exigua neonates respectively. Survival larvae and dead ones were counted on the 3rd,the 7th,the 10th,the 16th and the 19th day; meanwhile,the pupae amount was recorded,and the pupae weight was measured at the 24th h after pupation. [Result] The survival curves,pupation rates and pupae weights of S. exigua feeding on different tissues of transgenic cotton were not significantly different from those of S. exigua feeding on the corresponding tissues of conventional cotton; pupation rate of S. exigua feeding on different leaves of the same cotton variety were not significantly different from each other,but all higher than that of S. exigua feeding on the flowers of that cotton; and there were no differences among pupation weights of S. exigua feeding on different leaves or flowers of the same cotton variety. [Conclusion] Transgenic cotton showed weak resistance to S. exigua. Hence,in the transgenic cotton fields,more attention should be paid to occurrence trend of S. exigua and its control.

Screening of drought resistance indices and evaluation of drought resistance in cotton(Gossypium hirsutum L.)

Sixteen cotton cultivars widely planted in China were sowed under five different drought concentrations(0,2.5,5,7.5,and 10%)using PEG6000 to screen the indices of drought resistance identification and explore the drought resistance of different cotton cultivars.Eighteen physiological indices including root,stem,and leaf water contents(RWC,SWC,and LWC),net photosynthetic rate(Pn),the maximum photochemical quantum yield(Fv/Fm),the actual photochemical quantum yield(ΦPSII),non-photochemical quenching coefficient(NPQ),leaf water potential(LWP),osmotic potential(Ψs),leaf relative conductivity(REC),leaf proline content(Pro),leaf and root soluble protein contents(LSPC and RSPC),leaf and root malondialdehyde(MDA)contents(LMDA and RMDA),root superoxide dismutase,peroxidase,and catalase activities(RSOD,RPOD,and RCAT)were measured.Results indicated the 18 physiological indices can be converted into five or six independent comprehensive indices by principal component analysis,and nine typical indices(Fv/Fm,SWC,LWP,Pro,LMDA,RSPC,RMDA,RSOD,and RCAT)screened out by a stepwise regression method could be utilized to evaluate the drought resistance.Moreover,the 16 cotton cultivars were divided into four types:drought sensitive,drought weak sensitive,moderate drought resistant,and drought resistant types.The resistance ability of two selected cotton cultivars(drought resistant cultivar,Dexiamian 1;drought sensitive cultivar,Yuzaomian 9110)with contrasting drought sensitivities were further verified by pot experiment.Results showed that the responses of final cotton biomass,yield,and yield composition to drought were significantly different between the two cultivars.In conclusion,drought resistant cultivar Dexiamian 1 and drought sensitive cultivar Yuzaomian 9110 were screened through hydroponics experiment,which can be used as ideal experimental materials to study the mechanism of different cotton cultivars with contrasting drought sensitivities in response to drought stress.

Quantitative Trait Loci for Resistance Against Fusarium Wilt Based on Three Cotton F_2 Populations

Fusarium wilt （FW） is one of the most common cotton diseases in the world. Identification of QTLs conferring resistance to FW is key for the incorporation of resistance genes into elite cultivars. Two intraspecific （cross between Gossypium hirsuturn L.） and one interspecific （cross between Gossypium hirsutum L. and Gossypium bardence L.） F2 populations were constructed by using a highly resistant cultivar and crossing it to a susceptible cultivar with 154, 79, and 148 offsprings, respectively. Simple sequence repeats （SSR） were used to screen genomic regions closely linked to FW resistance. The results showed that five QTLs associated with FW resistance were detected in two intraspecific populations using a composite interval mapping method under four different conditions. Four of these loci located on Chr. 2/Chr. 17 neighboring markers JESPR304 or CIR305 which explained 13.1 to 45.9% of the phenotypic effect. Furthermore, JESPR304 and CIR305 were previously testified and found to be tightly linked. It is possible that these four QTLs detected under different conditions were the same resistance QTL/gene. We consider that there is the possibility of a major FW resistant gene in intraspecific populations. In the interspecific mapping populations two QTLs were detected on Chr. 9 and Chr. 12/26 which explained great phenotypic variance of 49.4 and 45.7%. As the location of QTLs for FW resistance among the intraspecific and the interspecfic populations were totally different, it is suggested that there may be different resistance mechanisms between G. bardence L. and G. hursutum L. Thus, the present research provides an opportunity to understand the genetic control of resistance to FW in Gossypium hirsutum and Gossypium bardence and to conduct MAS in breeding programs to develop FW resistant cultivars.

Pyramiding Breeding by Marker-Assisted Recurrent Selection in Upland Cotton: Selected Effects on Resistance to Helicoverpa armigera

The coincidence rates were more than 96% among the instar-weighted average of bioassaysin the lab, the percentage of resistance to Km in the field and the percentage of plantscontaining Bt gene. So, the performance of resistance to Km in the field can be used torepresent the transgenic Bt gene for selecting the resistance to bollworm. The instar-weighted averages were 30.585, 24.182, 16.615, 10.601, 10.123, 7.440 and 7.215 for theC0, P1, M1, M2, MP1, P2 and MP2 populations, respectively. The variance analysisindicated that the instar-weighted average in C0 was greatly significantly higher thanthat in all other populations, i.e., the performance of resistance to bollworm in C0 washighly significantly lower than all other populations. And the resistance in P1 wasgreatly lower than that of M1, M2, MP1, P2 and MP2, and M1 greatly lower than that of M2,MP1, P2 and MP2. There were no significant differences among M2, MP1, P2 and MP2. Withinthe populations of the first cycle selection, MP1 and M1 were greatly significantlyhigher than P1, and MP1 significantly higher than M1. The populations of the second cycleselection were significantly higher than their initial population M1, but no significantdifference among them. The boll size, seed index, the percent of the first harvest yield,fiber length, strength and elongation of the resistant plants to bollworm were significantlylower than that of sensitive plants to bollworm. And the yield of seed and lint cottonof the resistant plant to bollworm were lower than that of the sensitive to bollworm, butno significant difference between them. The boll numbers per plant, lint percent andmicronaire of the resistant plants to bollworm were significantly higher than that of thesensitive plant to bollworm.

Advances in Cotton Breeding for Resistance to Fusarium and Verticillium Wilt in the Last Fifty Years in China

This review summarizes the main advances in cotton breeding for resistance to fusarium and verticillium wilt in the last fifty years in China. The topics discussed include main achievements, experiences, problems and countermeasures for solving.

Succinate dehydrogenase SDH1–1 positively regulates cotton resistance to Verticillium dahliae through a salicylic acid pathway

Background:Verticillium wilt,caused by the soil-borne fungus of Verticillium dahliae Kleb.,is one of the most devastating diseases of cotton.The complex mechanism underlying cotton resistance to Verticillium wilt remains uncharacterized.Identifying an endogenous resistance gene may be helpful to control this disease.Previous studies revealed that succinate dehydrogenase(SDH)is involved in reactive oxygen species(ROS)-induced stress signaling pathway that is likely to be triggered by salicylic acid(SA).Here,through the metabolomics and differential expression analyses in wilt-inoculated cotton(Gossypium hirsutum),we noticed that Gh SDH1–1 gene in cotton may play an important role in the resistance to V.dahlia.Then we reported Gh SDH1–1 gene and its functional analysis in relation to the resistance of cotton to V.dahliae.Results:The Gh SDH1–1 gene in cotton root was significantly up-regulated after V.dahlia inoculation,and its expression level peaked at 12 and 24 h post-infection.SA can also induce the up-regulation of Gh SDH1–1.Additionally,the functional analysis showed that Gh SDH1–1-silenced cotton was more susceptible to V.dahliae than the control because of the significant decrease in abundance of immune-related molecules and severe damage to the SA-signaling pathway.In Arabidopsis thaliana,high expression of Gh SDH1–1 conferred high resistance to V.dahliae.Arabidopsis that overexpressed Gh SDH1–1 had higher resistance to V.dahliae infection compared with the wild-type.Conclusions:Our findings provide new insights into the role of Gh SDH1–1;it positively regulates cotton resistance to Verticillium wilt.The regulatory mechanism of Gh SDH1–1 is closely related to SA-related signaling pathway.

Insect resistance management in Bacillus thuringiensis cotton by MGPS(multiple genes pyramiding and silencing)

The introduction of Bacillus thuringiensis(Bt)cotton has reduced the burden of pests without harming the environment and human health.However,the efficacy of Bt cotton has decreased due to field-evolved resistance in insect pests over time.In this review,we have discussed various factors that facilitate the evolution of resistance in cotton pests.Currently,different strategies like pyramided cotton expressing two or more distinct Bt toxin genes,refuge strategy,releasing of sterile insects,and gene silencing by RNAi are being used to control insect pests.Pyramided cotton has shown resistance against different cotton pests.The multiple genes pyramiding and silencing(MGPS)approach has been proposed for the management of cotton pests.The genome information of cotton pests is necessary for the development of MGPS-based cotton.The expression cassettes against various essential genes involved in defense,detoxification,digestion,and development of cotton pests will successfully obtain favorable agronomic characters for crop protection and production.The MGPS involves the construction of transformable artificial chromosomes,that can express multiple distinct Bt toxins and RNAi to knockdown various essential target genes to control pests.The evolution of resistance in cotton pests will be delayed or blocked by the synergistic action of high dose of Bt toxins and RNAi as well as compliance of refuge requirement.

Differential Expression of Salinity Resistance Gene on Cotton

Salinity resistance and differential gene expression associated with salinity in cotton germplasm were studied,because of the large scale area of salinity in China,and its significant negative effects

Test on Wilt-Resistance of Simple Hybrides F7 and Beckross Hybrid Lines F6 B1 of <i>G. hirsutum</i>L. of Cotton to New Virulent Isolates of Fungus <i>Fusarium verticillioides</i>and <i>Verticillium dahliae</i>

The article presents the results of studies on the resistance of hybrid cotton lines to a new virulent isolate (strain) of the fungus Fusarium verticillioides upon inoculation of the host plant. Based on the studies, it was found that the complex genotypic resistance of the studied lines, when the host plants are inoculated with isolates of -100 V. dahliae Kleb fungus and 103 Fusarium verticillioides fungi, depends on the degree of resistance of the parental forms and their combination ability.

Evaluation on Diseases Resistance of Cotton Material and Its Utilization

Fusarium wilt and Verticillium wilt are important worldwide fungal diseases on cotton that cause damage to yield and quality.The pathogens survive in soil as microsclerotia for many years,and

Transgenic Cotton and Disease Resistance Genes

Success in conventional breeding for resistance to mycotoxin-producing or other phytopathogenic fungi is dependent on the availability of resistance gene(s) in the germplasm.Even when it is available,breeding for disease-resistant crops is very time consuming,especially in perennial crops such