Knockdown of the atypical protein kinase genes GhABC1K2-A05 and GhABC1K12-A07 make cotton more sensitive to salt and PEG stress

Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotton(Gossypium spp.).Here,we identified 40 ABC1Ks in upland cotton(Gossypium hirsutum L.)and found that the Gh ABC1Ks were unevenly distributed across 17 chromosomes.The GhABC1K family members included 35 paralogous gene pairs and were expanded by segmental duplication.The GhABC1K promoter sequences contained diverse cis-acting regulatory elements relevant to hormone or stress responses.The qRT-PCR results revealed that most Gh ABC1Ks were upregulated by exposure to different stresses.Gh ABC1K2-A05 and Gh ABC1K12-A07 expression levels were upregulated by at least three stress treatments.These genes were further functionally characterized by virus-induced gene silencing(VIGS).Compared with the controls,the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced cotton lines exhibited higher malondialdehyde(MDA)contents,lower catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)activities and reduced chlorophyll and soluble sugar contents under NaCl and PEG stress.In addition,the expression levels of six stress marker genes(Gh DREB2A,Gh SOS1,Gh CIPK6,Gh SOS2,Gh WRKY33,and Gh RD29A)were significantly downregulated after stress in the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced lines.The results indicate that knockdown of Gh ABC1K2-A05 and Gh ABC1K12-A07 make cotton more sensitive to salt and PEG stress.These findings can provide valuable information for intensive studies of Gh ABC1Ks in the responses and resistance of cotton to abiotic stresses.

GbLMI1 over-expression improves cotton aboveground vegetative growth

Leaves are the main organ for photosynthesis and organic synthesis in cotton.Leaf shape has important effects on photosynthetic efficiency and canopy formation,thereby affecting cotton yield.Previous studies have shown that LMI1(LATE MERISTEM IDENTITY1)is the main gene regulating leaf shape.In this study,the LMI1 gene was inserted into the 35S promoter expression vector,and cotton plants overexpressing LMI1(OE)were obtained through genetic transformation.Statistical analysis of the biological traits of the T_(1) and T_(2) populations showed that compared to the wild type(WT),OE plants had significantly larger leaves,thicker stems and significantly greater dry weight.Furthermore,plant sections of the main vein and petiole showed that the numbers of cells in those tissues of OE plants were significantly greater.In addition,RNA-seq analysis revealed the differential expression of genes related to gibberellin synthesis and NAC gene family(genes containing the NAC domain)between the OE and WT plants,suggesting that LMI1 is involved in secondary wall formation and cell proliferation,which promotes stem thickening.Moreover,Gene Ontology(GO)analysis revealed enrichment in the terms of calcium ion binding,and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis showed enrichment in the terms of fatty acid degradation,phosphatidylinositol signal transduction system,and c AMP(cyclic adenosine monophosphate)signal pathway.These results suggested that LMI1 OE plants are responsive to gibberellin hormone signals,and have altered messenger signals(c AMP,Ca^(2+))which amplify this function,to promote stronger aboveground vegetative growth.This study found the LMI1 greatly increased the vegetative growth in cotton,which is the basic requirement for higher yield.

Apical meristem transcriptome analysis identifies a role for the blue light receptor gene GhFKF1 in cotton architecture development

Cotton architecture is determined by the differentiation fate transition of axillary meristem(AM),and influences cotton yield and the efficiency of mechanized harvesting.We observed that the initiation of flowering primordium was earlier in early-maturing than that in late-maturing cultivars during the differentiation and development of AM.The RNA-Seq and expression level analyses showed that genes FLAVIN BINDING,KELCH REPEAT,F-BOX1(GhFKF1),and GIGANTEA(GhGI)were in response to circadian rhythms,and involved in the regulation of cotton flowering.The gene structure,predicted protein structure,and motif content analyses showed that in Arabidopsis,cotton,rapseed,and soybean,proteins GhFKF1 and GhGI were functionally conserved and share evolutionary origins.Compared to the wild type,in GhFKF1 mutants that were created by the CRISPR/Cas9 system,the initiation of branch primordium was inhibited.Conversely,the knocking out of GhGI increased the number of AM differentiating into flower primordium,and there were much more lateral branch differentiation and development.Besides,we investigated that proteins GhFKF1 and GhGI can interact with each other.These results suggest that GhFKF1 and GhGI are key regulators of cotton architecture development,and may collaborate to regulate the differentiation fate transition of AM,ultimately influencing plant architecture.We describe a strategy for using the CRISPR/Cas9 system to increase cotton adaptation and productivity by optimizing plant architecture.

The BEL1-like transcription factor GhBLH5-A05 participates in cotton response to drought stress

Drought stress impairs crop growth and development.BEL1-like family transcription factors may be involved in plant response to drought stress,but little is known of the molecular mechanism by which these proteins regulate plant response and defense to drought stress.Here we show that the BEL1-like transcription factor GhBLH5-A05 functions in cotton(Gossypium hirsutum)response and defense to drought stress.Expression of GhBLH5-A05 in cotton was induced by drought stress.Overexpression of GhBLH5-A05 in both Arabidopsis and cotton increased drought tolerance,whereas silencing GhBLH5-A05 in cotton resulted in elevated sensitivity to drought stress.GhBLH5-A05 binds to cis elements in the promoters of GhRD20-A09 and GhDREB2C-D05 to activate the expression of these genes.GhBLH5-A05 interacted with the KNOX transcription factor GhKNAT6-A03.Co-expression of GhBLH5-A05 and GhKNAT6-A03 increased the transcription of GhRD20-A09 and GhDREB2C-D05.We conclude that GhBLH5-A05 acts as a regulatory factor with GhKNAT6-A03 functioning in cotton response to drought stress by activating the expression of the drought-responsive genes GhRD20-A09 and GhDREB2C-D05.

A bZIP transcription factor GhVIP1 increased drought tolerance in upland cotton

Background Cotton is extremely affected by severe natural stresses.Drought is one of the most serious abiotic stress that adversely influences cotton growth,productivity,and fiber quality.Previous studies indicate that basic leucinezipper(bZIP)transcription factors are involved in the response of plants to various stresses.However,the molecular function and regulatory mechanism of GhVIP1 in response to drought stress are still unknown.Results In this research,GhVIP1 was cloned from a drought-tolerant variety.Expression of GhVIP1 was up-regulated in response to multiple abiotic stresses,especially under drought stress.And GhVIP1 was highly expressed in the root,stem,and 10 days post-anthesis ovule.Inhibiting the expression of GhVIP1 in cotton using the virus-induced gene silencing method resulted in higher electrical conductivity in leaves,but lower water content under drought stress compared with the WT plant.Overexpression of GhVIP1 in Arabidopsis enhanced plant drought tolerance through increasing the seed germination rate and improving the development of root.The exogenous expression of GhVIP1 up-regulated the transcription of genes associated with drought response and proline biosynthesis during drought stress in Arabidopsis.Conclusion In summary,these results indicated that GhVIP1 played a positive role in plants’response to drought stress.The use of GhVIP1 via modern biotechnology might facilitate the improvement of drought tolerance in cotton cultivars.

Cloning and Expression Analysis of a Brassinosteroid Biosynthetic Enzyme Gene,GhDWF1,from Cotton (Gossypium hirsuturm L.)

Brassinosteroids （BRs） are an important class of plant steroidal hormones that are essential in a wide variety of physiological processes. To determine the effects of BRs on the development of cotton fibers, through screening cotton fiber EST database and contigging the candidate ESTs, a key gene （GhDWF1） involved in the upstream biosynthetic pathway of BRs was cloned from developing fibers of upland cotton （Gossypium hirsutum L.） cv. Xuzhou 142. The full length of the cloned cDNA is 1 849 bp, including a 37 bp 5＇-untranslated region, an ORF of 1 692 bp, and a 120 bp 3＇-untranslated region. The cDNA encodes a polypeptide of 563 amino acid residues with a predicted molecular mass of 65 kD. The deduced amino acid sequence has high homology with the BR biosynthetic enzyme, DWARF1/DIMINUTO, from rice, maize, pea, tomato, and Arabidopsis. Furthermore, the typical conserved structures, such as the transmembrane domain, the FAD- dependent oxidase domain, and the FAD-binding site, are present in the GhDWF1 protein. The Southern blot indicated that the GhDWF1 gene is a single copy in upland cotton genome. RT-PCR analysis revealed that the highest level of GhDWF1 expression was detected in 0 DPA （day post anthesis） ovule （with fibers） while the lowest level was observed in cotyledon. The GhDWF1 gene presents high expression levels in root, young stem, and fiber, especially, at the fiber developmental stage of secondary cell wall accumulation. Moreover, the expression level was higher in ovules （with fibers） of wildtype （Xuzhou 142） than in ovules of fuzzless-lintless mutant at the same developmental stages （0 and 4 DPA）. The results suggest that the GhDWF1 gene plays a crucial role in fiber development.

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.

In-depth analysis of Bt cotton adoption:farmers'opinions,genetic landscape,and varied perspectives——a case study from Pakistan

Background Bt technology has played significant role in controlling bollworms and increasing cotton yield in earlier days of its introduction,a subsequent decline in yield became apparent over time.This decline may be attributed to various environmental factors,pest dynamics,or combination of both.Therefore,the present biophysical survey and questionnaire were designed to evaluate the impact of Bt cotton on bollworms management and its effect on reducing spray costs,targeting farmers with varied landholdings and educational backgrounds.Additionally,data on farmers'cultivated varieties and the prevalence of bollworms and sucking insects in their fields were recorded.Subsequently,about eleven thousand cotton samples from farmer fields were tested for Cry1Ac,Cry2Ab and Vip3A genes by strip test.Results In this analysis,83% of the farmers planting approved varieties believe that Bt technology control bollworms,while 17% hold contradictory views.Similarly,among farmers cultivating unapproved varieties,77% agree on effectiveness of Bt technology against bollworms,while 23% disagree.On the other hand,67% of farmers planting approved varieties believe that Bt technology does not reduce spray costs,while 33% agree with the effectiveness.Similarly,78% of farmers cultivating unapproved varieties express doubt regarding its role to reduce spray costs,while 22% are in favour of this notion.Differences in opinions on the effectiveness of Bt cotton in controlling bollworms and reducing spray cost between farmers planting unapproved and approved varieties may stem from several factors.One major cause is the heavy infestation of sucking insects,which is probably due to the narrow genetic variation of the cultivated varieties.Additionally,the widespread cultivation of unapproved varieties(21.67%)is also an important factor to cause different opinions on the effectiveness of Bt cotton.Conclusion Based on our findings,we propose that the ineffective control of pests on cotton crop may be attributed to large scale cultivation of unapproved varieties and non-inclusion of double and triple transgene technologies in country's sowing plan.On the basis of our findings,we suggest cotton breeders,regulatory bodies and legislative bodies to discourage the cultivation of unapproved varieties and impure seed.Moreover,the adoption of double and triple Bt genes in cottons with a broad genetic variation could facilitate the revival of the cotton industry,and presenting a promising way forward.

Development of New Interspecific Monosomic F_1 Substitution Stocks for Investigations in Cotton Genome

As it is known a complete coverage of cotton(Gossypium hirsutum L.) genome with hypoaneuploids is not still realized.Therefore the detection of new types of aneuploids especially in different cytogenetic collections is very useful.In Uzbekistan,long-term investigations towards development

Genotypic Resistance of F_1 Cotton Hybrids by Inoculation with Different Virulent Isolates of the Fungus Verticillium Dahliae Klebahn

The plant pathogen Verticillium dahliae causes severe cotton losses in Uzbekistan. To create cotton varieties that are resistant to the more virulent races of V.dahliae we wanted to determine

U.S.Cotton Industry Continuing Dialogue on Quality with No.1 Customer

MEMPHIS-A National Cotton Council delegation traveling to Beijing on June 19-30 will make a number of visits with key Chinese officials to:1)provide updates on the U.S. cotton industry’s efforts to improve its cotton quality and 2) ascertain its Chinese textile customers’needs.

Cotton Plants Transformed with the Activated Chimeric Cry1Ac and API-B Genes

A chimeric gene, Bt29K, composed of coding sequences of activated Cry1Ac insecticidal protein and an endoplasm reticulum-retarding signal peptide, was synthesized. A plant expression vector containing two expression cassettes for the Bt29K and API-B genes was constructed. These two insect-resistant genes were transferred into two cotton ( Gossypium hirsutum L.) varieties ( or lines) via Agrobacterium-mediated transformation and nine homozygous transgenic cotton lines showing a mortality of 90.0% - 99.7% to cotton ballworm (Heliothis armigera) larvae and good agronomic traits were selected through six generations. Molecular biology analysis revealed that one or two copies of the insecticidal protein genes were integrated into the transgenic cotton genome and activated Cry1Ac and API-B protein expression was at a level of 0.17% and 0.09% of the total soluble protein in the transgenic cotton leaves, respectively. Comparison of the insect-resistance of the homozygous lines expressing the activated chimeric Cry1Ac and API-B with that expressing Cry1Ac only revealed that the insect-resistance of the former is apparently higher than the latter. These results also indicate that the strategy to construct a plant expression vector expressing two different insect-resistant genes reported here is reasonable.

Obtaining High Pest_resistant Transgenic Upland Cotton Cultivars Carrying cry1Ac3 Gene Driven by Chimeric OM Promoter

Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.

Cloning of Cotton Delta-12 Oleate Desaturase Gene FAD2-1 and Construction of Its ihpRNA and amiRNA Interference Vectors

Delta-12 oleate desaturase gene （FAD2-1） which converts oleic acid into linoleic acid, is the key enzyme determining the fatty acid composition of cottonseed oil. By employing RT-PCR method, full length cDNA of cotton delta-12 oleate desat- urase gene GhFAD2-1 containing an open reading frame of 1 158 bp was cloned for constructing RNAi vector. A 515 bp long specific fragment of this gene was se- lected for constructing ihpRNA vector under the control of a seed-specific promoter NAPIN, named pFGC1008-NAPIN-FAD2-1; meanwhile miRNA gene-silencing vector pCAMBIA1302-amiRNA-FAD2-1 targeting GhFAD2-1 was also constructed.

Cotton BLH1 and KNOX6 antagonistically modulate fiber elongation via regulation of linolenic acid biosynthesis

BEL1-LIKE HOMEODOMAIN(BLH)proteins are known to function in various plant developmental processes.However,the role of BLHs in regulating plant cell elongation is still unknown.Here,we identify a BLH gene,GhBLH1,that positively regulates fiber cell elongation.Combined transcriptomic and biochemical analyses reveal that GhBLH1 enhances linolenic acid accumulation to promote cotton fiber cell elongation by activating the transcription of GhFAD7A-1 via binding of the POX domain of GhBLH1 to the TGGA cis-element in the GhFAD7A-1 promoter.Knockout of GhFAD7A-1 in cotton significantly reduces fiber length,whereas overexpression of GhFAD7A-1 results in longer fibers.The K2 domain of GhKNOX6 directly interacts with the POX domain of GhBLH1 to form a functional heterodimer,which interferes with the transcriptional activation of GhFAD7A-1 via the POX domain of GhBLH1.Overexpression of GhKNOX6 leads to a significant reduction in cotton fiber length,whereas knockout of GhKNOX6 results in longer cotton fibers.An examination of the hybrid progeny of GhBLH1 and GhKNOX6 transgenic cotton lines provides evidence that GhKNOX6 negatively regulates GhBLH1-mediated cotton fiber elongation.Our results show that the interplay between GhBLH1 and GhKNOX6 modulates regulation of linolenic acid synthesis and thus contributes to plant cell elongation.

The bHLH transcription factor GhPAS1 mediates BR signaling to regulate plant development and architecture in cotton

Cotton (Gossypium spp.) is the most important natural textile fiber crop in the world.The ideal plant architecture of cotton is suitable for mechanical harvesting and productivity in modern agricultural production.However,cotton genes regulating plant development and architecture have not been fully identified.We identified a basic helix-loop-helix (b HLH) transcription factor,GhPAS1 (PAGODA1 SUPPRESSOR1) in G.hirsutum (Upland cotton).GhPAS1 was located in the nucleus and showed a strong transcription activation effect.Tissue-specific expression patterns showed that GhPAS1 was highly expressed in floral organs,followed by high expression in early stages of ovule development and rapid fiber elongation.GhPAS1 overexpression in Arabidopsis and BRZ (brassinazole,BR biosynthesis inhibitor) treatment indicated that GhPAS1 positively regulates and responds to the BR (brassinosteroid) signaling pathway and promotes cell elongation.GhPAS1 overexpression in Arabidopsis mediated plant development in addition to increasing plant biomass.Virus-induced gene silencing of GhPAS1 indicated that down-regulation of GhPAS1 inhibited cotton growth and development,as plant height,fruit branch length,and boll size of silenced plants were lower than in control plants.Fiber length and seed yield were also lower in silenced plants.We conclude that GhPAS1,a b HLH transcription factor,regulates plant development and architecture in cotton.These findings may help breeders and researchers develop cotton cultivars with desirable agronomic characteristics.

Genetic variation in LBL1 contributes to depth of leaf blades lobes between cotton subspecies, Gossypium barbadense and Gossypium hirsutum

Leaf is a essential part of the plants for photosynthetic activities which mainly economize the resources for boll heath. Significant variations of leaf shapes across the Gossypium sp. considerably influence the infiltration of sunlight for photosynthesis. To understand the genetic variants and molecular processes underlying for cotton leaf shape, we used F2 population derived from upland cotton genotype P30A （shallow-lobed leaf） and sea-island cotton genotype ISR （deep-lobed leaf） to map leaf deep lobed phenotype controlling genes LBL1 and LBL2. Genetic analysis and localization results have unmasked the position and interaction between both loci of LBL1 and LBL2, and revealed the co-dominance impact of the genes in regulating depth of leaf blades lobes in cotton. LBL1 had been described as a main gene and member of transcription factor family leucine zipper （HD-ZIPI） from a class I homologous domain factor Gorai.OO2G244000. The qRT-PCR results elaborated the continuous change in expression level of LBL1 at different growth stages and leaf parts of cotton. Higher expression level was observed in mature large leaves followed by medium and young leaves respectively. For further confirmation, plants were tested from hormonal induction treatments, which explained that LBL 1 expression was influenced by hormonal signaling. Moreover, the highest expression level was detected in brassinolides （BR） treatment as compared to other hormones, and this hormone plays an important role in the process of leaf blade lobed formation.

Effect of 1-MCP on Gas Exchange and Carbohydrate Concentrations of the Cotton Flower and Subtending Leaf under Water-Deficit Stress

Ethylene is an endogenous plant hormone that increases under adverse environmental conditions, resulting in leaf and fruit abscission and ultimately yield reduction. In cotton, however, the effects of water-deficit stress on ethylene production have been uncertain. In this study it was hypothesized that application of an ethylene inhibitor 1-Methylcyclo- propene (1-MCP) would prevent ethylene production and result in alleviation of water-deficit stress consequences on the physiology and metabolism of the cotton flower and subtending leaf. To test this hypothesis, growth chamber experiments were conducted in 2009-2010 with treatments consisting of (C) untreated well-watered control, (C + 1MCP) well-watered plus 1-MCP, (WS) untreated water-stressed control, and (WS + 1MCP) water-stressed plus 1-MCP. The plants were subjected to two consecutive drying cycles during flowering, approximately 8 weeks after planting, and 1-MCP was foliar applied at a rate of 10g. ai/ha at the beginning of each drying cycle. The results showed that 1-MCP application had no significant effect on gas exchange functions and did not prevent reductions from water stress in leaf photosynthesis, respiration and stomatal conductance. However, application of 1-MCP resulted in a decrease in sucrose content of water-stressed pistils compared to the control indicating that 1-MCP has the potential to interfere in carbohydrate metabolism of reproductive units.

Cotton GhBRC1 regulates branching,flowering,and growth by integrating multiple hormone pathways

Cotton architecture is partly determined by shoot branching and flowering patterns.Gh BRC1 was previously identified by RNA-seq analysis of nulliplex-branching and normal-branching cotton.However,the roles of Gh BRC1 in cotton remain unclear.In the present study,investigations of nuclear localization and transcriptional activity indicated that Gh BRC1 has characteristics typical of transcription factors.Gene expression analysis showed that Gh BRC1 was highly expressed in axillary buds but displayed different expression patterns between the two branching types.Overexpression of Gh BRC1 in Arabidopsis significantly inhibited the number of branches and promoted flowering.In contrast,silencing Gh BRC1 in cotton significantly promoted seedling growth.Gh BRC1 was induced by multiple hormones,including strigolactones,which promoted seedling growth and seed germination of Arabidopsis plants overexpressing Gh BRC1.Consistent with these findings,RNA-seq analysis of virus-induced gene silencing treated cotton revealed that a large number of genes were differentially expressed between Gh BRC1-silenced and control plants,and these genes were significantly enriched in plant hormone signalling pathways.Together,our data indicates that Gh BRC1 regulates plant branching and flowering through multiple regulatory pathways,especially those regulating plant hormones,with functions partly differing from those of Arabidopsis BRC1.These results provide insights into the molecular mechanisms controlling plant architecture,which is important for breeding cotton with ideal plant architecture and high yield.

Does cotton bollworm show cross-resistance to the Bacillus thuringiensis toxins Cry1Ac and Cry2Ab? A mini review

Since 1996, transgenic Bacillus thuringiensis(Bt) cotton has been commercially grown in numerous countries in an effort to stem the losses caused by key lepidopteran pests. However, the development of pest resistance to Bt toxins has jeopardized the continued utilization of Bt cotton. As a strategy designed to circumvent the development of resistance, Bt cotton varieties expressing two or more toxins targeting the same pest have been introduced. Nevertheless, from the perspective of long-term planting of Bt cotton, the potential risk of cross-resistance to these Bt toxins is a threat that cannot be ignored. In this paper, we review current research(including that based on the analysis of protein binding sites and resistance genes) on the resistance of cotton bollworm(Helicoverpa armigera) to the Bt toxins Cry1 Ac and Cry2 Ab and the interrelationship between these toxins. On the basis of existing evidence, we assume that the actions of Cry1 Ac and Cry2 Ab against cotton bollworm are not completely independent, and then propose the "resistance-associated gene mutation potential hypothesis". Although the mechanisms underlying the resistance of pests to Bt toxins are yet to be comprehensively elucidated, this hypothesis could undoubtedly have important implications for adopting "pyramid" strategy in the future. Further research is recommended to devise strategies to retard the development of H. armigera resistance to Bt cotton, either using different Bt toxins or their various combinations.