Detection of ARV-Resistant Mutants in HIV-1-Infected Individuals in a Context of Systematic Switching to an Association Based on Dolutegravir in Abidjan, Côte d’Ivoire

The emergence of antiretroviral resistance mutations represents a major threat to the achievement of national and global goals for the elimination of HIV-1 infection. The global strategy in 2019 in Cte d'Ivoire is a new national policy for the management of people living with HIV with the administration of dolutegravir (DTG)-based fixed-dose combination. The aim of our study was to evaluate HIV-1 resistance to antiretrovirals (ARVs) in infected adult subjects in Cte d’Ivoire in the context of a systematic switch to a DTG-based combination. Between February 2022 and October 2023, a cross-sectional survey with random sampling was conducted in 06 services caring for people living with HIV. A total of 139 participants were included in the study. Adults with a viral load ≥ 1000 copies/mL were tested for HIV-1 ARV resistance mutations. Molecular analyses were performed using protocol of ANRS-MIE (National Agency for Research on AIDS and emerging infectious diseases). The interpretation is performed by HIVGRAD (https://www.hiv-grade.de/cms/grade/). The frequencies of HIV-1 resistance to non-nucleotide reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NRTIs), integrase inhibitors (IINTs) and protease inhibitors (PIs) were 82%, 73%, 19% and 11% respectively. The main mutations observed in the different classes were K103N (45%), M184V (64%), E157Q (19%) and L10V/M46I/A71V/I54V (6%) respectively. This study reveals the emergence of resistance to DTG-based fixed-dose combinations, favored by high rates of resistance to NRTIs and NNRTIs. This finding underlines the need for enhanced viral load monitoring and HIV-1 genotyping tests to guide the choice of NRTIs for combination therapy. In addition, monitoring for mutations to second-generation NRTIs is essential, given the scale-up of DTG-based regimens currently underway in Cte d’Ivoire.

Mutant Selection Windows of Azalomycin F5a in Combination with Vitamin K3 against Methicillin-Resistant Staphylococcus aureus

Azalomycin F5a, a 36-membered macrocyclic lactone isolated from several streptomyces strains, presented remarkable anti-methicillin-resistant Staphylococcus aureus (MRSA) activities. To improve its anti-MRSA potential and to evaluate the probability of MRSA resistant to it before development, the anti-MRSA activities of azalomycin F5a in combination with vitamin K3 were first evaluated using checkerboard assay. Then the minimal concentration inhibiting colony formation by 99% (MIC99) and mutant prevention concentration (MPC) of azalomycin F5a alone and in combination with vitamin K3 against MRSA were determined using agar plates with linear antimicrobial concentration decrease. The fractional inhibitory concentration indexes (FICIs) of 0.25 - 0.50 showed the synergistic activity of azalomycin F5a in combination with vitamin K3. The mutant selection windows (MSWs, MIC99-MPC) of azalomycin F5a alone against MRSA tested were 2.07 - 6.40 μg/mL, and the MPCs of azalomycin F5a in combination with vitamin K3 against MRSA tested were 1.60 - 3.20 μg/mL. These indicated that the MPCs of azalomycin F5a in combination could drop down to below its MIC99 alone. According to the hypothesis of MSW, the narrower MSWs of azalomycin F5a alone, even closed MSWs in combination with vitamin K3, together with their synergistic anti-MRSA activities, indicated that azalomycin F5a had a good potential to develop as a new antimicrobial agent.

Responses of Drought-Resistant Mutant vem1 to Stress and Cloning of VEM1 Gene in Arabidopsis

[ Objective] This study aimed to investigate the responses of drought-resistant mutant veml to stress and clone VEM1 gene in Arabidopsis. [ Method] A drought-resistant mutant veml was isolated from the Arabidops/s mutant pool. The germination rates of wild-type （WT） and mutant veml were detected to investigate the responses of mutant veml to mannitol, NaCl and ABA stress. [ Result] The mutant veml was resistant to mannitol and NaC1 stress but sensitive to ABA stress. VEM1 gene was cloned by Tail-PCR technology and sequenced. The sequencing result was submitted to NCBI for sequence alignment and gene mapping using BLAST. Database analysis suggested that VEM1 gene was a transposable clement gene. [ Conclusion] This study laid the foundation for functional analysis of drought-resistant gene VEM1.

Chemical mutagenesis and soybean mutants potential for identification of novel genes conferring resistance to soybean cyst nematode

The resistance of soybean(Glycine max(L.) Merr.) to soybean cyst nematode(SCN, Heterodera glycines Ichinohe), which is a devastating pathogen in soybean production and causes a large quantity of annual yield loss worldwide, can shift during the long-term interaction and domestication. It is vital to identify more new resistance genetic sources for identification of novel genes underlying resistance to SCN for management of this pathogen. In the present study, first, two ethane methylsulfonate-mutagenesis soybean M2 populations of PI 437654, which shows a broad resistance to almost all of SCN races, and Zhonghuang 13, which is a soybean cultivar in China conferring strong resistance to lodging, were developed. Many types of morphological phenotypes such as four-and five-leaflet leaves were observed from these two soybean M2 populations. Second, 13 mutants were identified and confirmed to exhibit alteration of resistance to SCN race 4 through the forward genetic screening of 400 mutants of the PI 437654 M2 population, the rate of mutants with alteration of SCNinfection phenotype is 3.25%. Third, these identified mutants were further verified not to show any changes in the genomic sequences of the three known SCN-resistant genes, GmSHMT08, GmSNAP18 and GmSANP11, compared to the wildtype soybean; and all of them were still resistant to SCN race 3 similar to the wild-type soybean. Taken together, we can conclude that the 13 mutants identified in the present study carry the mutations of the new gene(s) which contribute(s) to the resistance to SCN race 4 in PI 437654 and can be potentially used as the genetic soybean sources to further identify the novel SCN-resistant gene(s).

Identification and Genetic Analysis of a Novel Rice Spotted-Leaf Mutant with Broad-Spectrum Resistance to Xanthomonas oryzae pv. oryzae

A spotted-leaf mutant of rice HM143 was isolated from an EMS-induced IR64 mutant bank. Brown lesions randomly distributed on leaf blades were observed about 3 wk after sowing. The symptom lasted for the whole plant growth duration. Histochemical analysis indicated that cell death occurred in and around the site of necrotic lesions accompanied with accumulation of hydrogen hyperoxide. Agronomic traits were largely similar to the wild type IR64 except seed setting rate and 1 000-grain weight which were significantly decreased in the mutant. Disease resistance of the mutant to multiple races of Xanthomonas oryzae pv. oryzae was significantly enhanced. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively termed splHM143. In addition, using molecular markers and 1023 mutant type individuals from an F2 segregating population derived from the cross HM143/R9308, the spotted-leaf gene was finally delimited to an interval of 149 kb between markers XX25 and ID40 on the long arm of chromosome 4. splHM143 is likely a novel rice spotted-leaf gene since no other similar genes have been identified near the chromosomal region.

Transferring Translucent Endosperm Mutant Gene Wx-mq and Rice Stripe Disease Resistance Gene Stv-bi by Marker-Assisted Selection in Rice (Oryza sativa)

A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-b＇ and a translucent endosperm mutant gene Wx-mq. From F2 generations, a sequence characterized amplified region （SCAR） marker tightly linked with Stv-b＇ and a cleaved amplified polymorphic sequence （CAPS） marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, Ning 9108, with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance. The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality, disease resistance and high yield in rice breeding.

Characterization of salt tolerance and Fusarium wilt resistance of a sweetpotato mutant

The variant LM1 was previously obtained using embryogenic cell suspension cultures of sweetpotato variety Lizixiang by gamma-ray induced mutation, and then its characteristics were stably inherited through six clonal generations, thus this mutant was named LM1. In this study, systematic characterization of salt tolerance and Fusarium wilt resistance were performed between Lizixiang and mutant LM1. LM1 exhibited significantly higher salt tolerance compared to Lizixiang. The content of proline and activities of superoxide dismutase（SOD） and photosynthesis were significantly increased, while malonaldehyde（MDA） and H_2O_2 contents were significantly decreased compared to that of Lizixiang under salt stress. The inoculation test with Fusarium wilt showed that its Fusarium wilt resistance was also improved. The lignin, total phenolic, jasmonic acid（JA） contents and SOD activity were significantly higher, while H_2O_2 content was significantly lower in LM1 than that in Lizixiang. The expression level of salt stress-responsive and disease resistance-related genes was significantly higher in LM1 than that in Lizixiang under salt and Fusarium wilt stresses, respectively. This result provides a novel and valuable material for improving the salt tolerance and Fusarium wilt resistance of sweetpotato.

Outcome of lamivudine-resistant hepatitis B virus is generally benign except in cirrhotics

AIM： We set to determine factors that determine clinical severity after the development of resistance.METHODS： Thirty-five Asian patients with genotypic lamivudine resistance were analyzed in three groups： 13/35 （37%） were non-cirrhotics with normal pre-treatment ALT （Group IA）, 12/35 （34%） were non-cirrhotics with elevated pre-treatment ALT （Group IB）, and 10/35 （29%） were cirrhotics （Group II）. Patients were followed for a median of 98 wk （range 26-220） after the emergence of genotypic resistance.RESULTS： Group IA patients tended to retain normal ALT. Group IB patients showed initial improvement of ALT with lamivudine but 9/12 patients （75%） developed abnormal ALT subsequently. On follow-up however, this persisted in only 33%. Group II patients also showed improvement while on treatment, but they deteriorated with the emergence of resistance with 30% death from decompensated liver disease. Pretreatment ALT levels and CPT score （in the cirrhotic group） were predictive of clinical resistance and correlated with peak ALT levels and CPT score.CONCLUSION： The phenotype of lamivudine-resistant HBV correlated with the pretreatment phenotype. The clinical course was generally benign in non-cirrhotics. However, cirrhotics had a high risk of progression and death （30%） with the development of lamivudine resistance.

Resistance to Bacterial Leaf Blight in a Somaclonal Rice Mutant HX-3 at Cellular Level

The interaction between rice host and its pathogen Xanthomonas oryzae pv. oryzae (Xoo) at cellular level was studied by using a resistant somaclonal mutant HX-3 and its susceptable donor Minghui 63. After inoculation with Xoo strain Zhe 173 (Chinese pathotype Ⅳ), the activity of superoxide dismutase (SOD) and peroxidase (POD) in the callus of Minghui 63 was increased dramatically, and the active oxygen(O2 ) was produced at a higher rate; Meanwhile, the callus grew slowly with the reduction of protein content Compared to the activity of SOD and POD, the production rate of Oa and the fresh weight in HX-3 callus varied little after the inoculation It could be proposed that there were great differences between the resistance of HX-3 and Mighui 63 at cellular level. There was no difference detected concerning resistance to bacterial leaf blight in HX-3 between the plant and the callus.

Characterization and mapping of a novel light-dependent lesion mimic mutant lmm6 in rice(Oryza sativa L.)

A novel rice lesion mimic mutant (LMM) was isolated from an ethane methyl sulfonate (EMS)-induced 02428 mutant bank. The mutant, tentatively designated as lmm6, develops necrotic lesions in the whole growth period along with changes in several important agronomic traits. We found that the initiation of the lesions was induced by light and cel death occurred in lmm6 accompanied with accumulation of reactive oxygen species (ROS). The lower chlorophyl content, soluble protein content and superoxide dismutase (SOD) activity, the higher malondialdehyde (MDA) content were detected in lmm6 than in the wild type (WT). Moreover, the observation by transmission electronic microscope (TEM) demonstrated that some organel es were damaged and the stroma lamel a of chloroplast was irregular and loose in mesophyl cel of lmm6. In addition, lmm6 was more resistant than WT to rice blast fungus Magnaporthe grisea infection, which was consistent with increased expression of four genes involved in the defense-related reaction. Genetic analysis showed that mutant trait of lmm6 is inherited as a monogenic recessive nuclear gene located on the long arm of chromosome 6. Using simple sequence repeat (SSR) markers, the target gene was ifnal y delimited to an interval of 80.8 kb between markers MM2359 and MM2370, containing 7 annotated genes. Taken together, our results provide the information to identify a new gene involved in rice lesion mimic, which wil be helpful in clarifying the mechanism of cel death and disease resistance in rice.

Efflux pump gene hefA of Helicobacter pylori plays an important role in multidrug resistance

AIM: To determine whether efflux systems contribute to multidrug resistance of H pylori. METHODS: A chloramphenicol-induced multidrug resistance model of six susceptible H pylori strains (5 isolates and H pylori NCTC11637) was developed. Multidrug-resistant (MDR) strains were selected and the minimal inhibitory concentration (MIC) of eryth-romycin, metronidazole, penicillin G, tetracycline, and ciprofloxacin in multidrug resistant strains and their parent strains was determined by agar dilution tests. The level of mRNA expression of hefA was assessed by fluorescence real-time quantitative PCR. A H pylori LZ1026 knockout mutant (ΔH pylori LZ1026) for (puta-tive) efflux protein was constructed by inserting the kanamycin resistance cassette from pEGFP-N2 into hefA, and its susceptibility profiles to 10 antibiotics were evaluated. RESULTS: The MIC of six multidrug-resistant strains (including 5 clinical isolates and H pylori NCTC11637) increased signifi cantly (≥ 4-fold) compared with their parent strains. The expression level of hefA gene was significantly higher in the MDR strains than in their parent strains (P = 0.033). A H pylori LZ1026 mutant was successfully constructed and the ΔH pylori LZ1026 was more susceptible to four of the 10 antibiotics. All the 20 strains displayed transcripts for hefA that con-fi rmed the in vitro expression of these genes.CONCLUSION: The efflux pump gene hefA plays an important role in multidrug resistance of H pylori.

Development of herbicide resistance genes and their application in rice

Rice is one of the most important food crops in the world.Weeds seriously affect the rice yield and grain quality.In recent years,there are tremendous progresses in the research and application of herbicideresistant genes in rice worldwide.This article reviews the working mechanisms of six herbicides(glyphosate,glufosinate,acetolactate synthase inhibitor herbicides,acetyl-Co A carboxylase inhibitor herbicides,hydroxyhenylpyruvate dioxygenase(HPPD)inhibitor herbicides and dinitroaniline herbicides),the resistance mutations of the corresponding herbicide-target genes,and the herbicide detoxification mechanisms by non-target genes.Examples are provided on herbicide-resistant rice materials obtained by transformation of exogenous resistance genes,by artificial mutagenesis and mutant screening,and by modifying the target genes through gene editing.This paper also introduces the current application of herbicide-resistant rice,points out problems that may be caused by utilization of herbicide resistant rice and solutions to the problems,and discusses the future prospects for the development of herbicideresistant rice.

Field identification of morphological and physiological traits in two special mutants with strong tolerance and high sensitivity to drought stress in upland rice(Oryza sativa L.)

The two mutants idr1-1 and 297-28, which were obtained from the radiation mutation of HD297 and IAPAR9, were used as experimental materials in this study for a 2-year(2012 and 2013) experiment about field drought resistance identification in Beijing, China. Key agronomic traits and water-related physiological indexes were observed and measured, including the leaf anti-dead level(LADL), days to heading, plant height, setting percentage, aboveground biomass, leaf water potential(LWP), net photosynthetic rate(Pn) and transpiration rate. The results showed that the mutant idr1-1 that was under drought stress(DS) conditions for 2 years had the highest LADL grades(1.3 and 2.0) among all the materials, and they were 2–3 grades stronger than the wild-type IAPAR9 with an average that was 21.4% higher for the setting percentage than the wild type. Compared with the IAPAR9 for the 2-year average delay in the days to heading and the reduction rates in the plant height, setting percentage, and aboveground biomass under DS compared with the well-watered(WW) treatment, idr1-1 showed 3.2% less delay and 19.1, 16.4, and 6.1% less reduction, respectively. The idr1-1 in the LWP always exhibited the highest performance among all the materials. The Pn of idr1-1 under severe and mild DS comparing with that under WW was slightly decreased and even slightly increased, respectively, leading to an average reduction rate of only 0.92%, which was 26.93% less than that of IAPAR9. Under the severe DS, idr1-1 still showed the highest value of 16.88 μmol CO2 m–2 s–1 among all the materials and was significantly higher than that of IAPAR9(11.66 μmol CO2 m–2 s–1). Furthermore, only idr1-1 had the increased and the highest transpiration rate values(7.6 and 6.04 mmol H2 O m–2 s–1) under both mild and severe DS compared with the values under WW, when the transpiration rate of all the other materials significantly decreased. By contrast, the 297-28 in terms of the LADL grade under DS was the lowest(7.0), and it was four grades weaker than its wildtype HD297 and even one grade weaker than the drought-sensitive paddy rice SN265. For the 2-year average reduction rates in aboveground biomass and plant heights under DS compared with those under the WW, 297-28 was 31.6 and 31.8% higher than HD297, respectively. Meanwhile, 297-28 showed the worst performance for the LWP, Pn, and transpiration rate. These results suggest that idr1-1 might be a superior drought tolerant mutant of upland rice found in China. It has a strong ability to maintain and even enhance leaf transpiration while maintaining a high plant water potential under DS, thus supporting a high Pn and alleviating the delay in agronomic trait development and yield loss effectively. 297-28 is a much more highly drought-sensitive mutant that is even more sensitive than paddy rice varieties. The two mutants could be used as drought tolerance controls for rice germplasm identification and the drought resistant mechanism studies in the future. idr1-1 is also suitable for breeding drought-tolerant and lodging-resistant high-yield rice varieties.

The antioxidant protein ZmPrx5 contributes resistance to maize stalk rot

Maize(Zea mays L.)stalk rot is a devastating disease worldwide,causing severe yield losses.Although previous studies have focused on the genetic dissection of maize resistance to stalk rot,the mechanisms of resistance remain largely unknown.We used a comparative proteomics approach to identify candidate proteins associated with stalk rot resistance.Statistical analyses revealed 763 proteins differentially accumulated between Fusarium graminearum and mock-inoculated plants.Among them,the antioxidant protein ZmPrx5,which was up-accumulated in diseased plants,was selected for further study.ZmPrx5 transcripts were present in root,stalk,leaf,ear,and reproductive tissues.The expression of ZmPrx5 in three inbred lines increased significantly upon F.graminearum infection.ZmPrx5 was localized in the cytoplasm.Compared to control plants,maize plants overexpressing ZmPrx5 showed increased resistance to F.graminearum infection,and ZmPrx5 mutant plants were more susceptible than wild-type plants.Defense-associated pathways including plant–pathogen interactions,phenylalanine metabolism,and benzoxazinoid and flavonoid biosynthesis were suppressed in ZmPrx5 homozygous mutant plants compared with wild-type plants.We suggest that ZmPrx5 positively regulates resistance against stalk rot in maize,likely through defense-oriented transcriptome reprogramming.These results lay a foundation for further research on the roles of Prx5 subfamily proteins in resistance to plant fungal diseases,and provide a potential genetic resource for breeding disease-resistance maize lines.

Reaction of Sesame (<i>Sesamum indicum</i>Linn.) Mutant Generations against Webworm, <i>Antigastra catalaunalis</i>Duponchel

Sesame (Sesamum indicum Linn.) (Pedaliaceae) is an important oilseed crop grown in many countries. Among the insect pests infesting sesame, the webworm, Antigastra catalaunalis Duponchel (Pyraustidae: Lepidoptera) is predominant throughout the crop period. For managing this insect, resistant sesame varieties with higher yield potential and better adaptability to varied locations are essentially needed. Keeping this in view and based on earlier work, three promising accessions viz., IVTS-2001-7, KMR-102 and TMV-3 were selected. To enhance resistance and/or yield traits, these three accessions in comparison with a susceptible check SVPR 1 were subjected to mutagenesis using gamma rays as physical mutagen;Ethyl Methane Sulphonate (EMS) and Diethyl Sulphate (DES) as chemical mutants. The first and generation mutants were evaluated under field conditions at Methikudi village, Cuddalore district, Tamil Nadu, Southern India during May, 2012-September, 2014. Webworm infestation was evaluated based on leaf, flower and capsule damage. Among the first mutant (M1) and second mutant (M2) generations, plants of the accessions namely IVTS 2001-7 and TMV-3 were rated as resistant and plants of SVPR-1 were highly susceptible to A. catalaunalis.

Multiple Resistance to Acetyl Coenzyme A Carboxylase and Acetolactate Synthase Inhibiting Herbicides in Tunisian Ryegrass Populations （Lolium rigidum）

The good understanding of the mechanisms of resistance to herbicides in weeds is a necessity to implement sustainable weed management strategies. Here, a study was conducted to characterize the molecular bases of resistance to acetyl coenzyme A carboxylase （ACCase） and acetolactate synthase （ALS） inhibiting herbicides in Lolium rigidum populations from Tunisia. Nine Lolium rigidum （ryegrass） populations collected in wheat fields from Northern Tunisia were investigated for their resistance to two ACCase-inhibiting herbicides and an ALS-inhibiting herbicide. All populations were tested in the greenhouse in pots using the commercial dose to determine resistance status. Survival plants were also tested for the presence of two ACCase （L 1781 and N2041） and two ALS （P197 and W574） mutant resistant alleles using molecular markers. Resistance to ACCase-inhibiting herbicides was found in all tested populations. Comparison of the results from herbicide sensitivity bioassays with genotyping indicated that more than 80% of the plants resistant to ACC-inhibiting herbicides would be resistant via increased herbicide metabolism. However, ALS-inhibiting herbicides are still more or less controlling ACCase resistant populations, so indicating that the selection process of resistance is ongoing. Target-site resistance appears to be the major mechanism for these early cases of ALS inhibitor resistance. This study reported the first case of resistance to ALS-inhibiting herbicides in ryegrass in Tunisia, and investigated the molecular bases of this resistance. It establishes the clear importance of non target-site resistance to ACCase- and/or ALS-inhibiting herbicides.

Construction and Verification of LuxS-negative Mutants of Streptococcus Mutans and the Effect of the Absence of LuxS Gene on the Acid Tolerance

Objective: To knock out the entire Luxs gene of Streptococcus mutans(S.mutans) UA159 strain via homologous recombination and construct a Luxs-deleted mutant strain of S. mutans. To study the difference between the acid resistance of S. mutans Ingbritt C international standard strain and the acid resistance of LuxS mutant strain. Methods: Two DNA fragments locating in the upper and downstream of Luxs gene were amplified and a erythromycin resistance gene of PJT10 between them were engineered into PUC19 plasmid for constructing the recombination plasmid pUCluxKO. Electrotransformation of S.mutans cells with pUCluxKO-mutant resulted in isolation of erythromycin resistant S. mutans transformants, which was identified by polymerase chain reaction, V.harveyi BB170 luminescence bioassay and sequencing analysis. Solutions of S. mutans standard strain and LuxS mutant strain with same density were made and cultured at pH 3.5 to 7.0 BHI liquid for the same period.Terminal growth situation was compared.Firstly acidized in pH 5.5 BHI liquid,the two strains were cultured at pH 3.0 BHI liquid. The acid tolerance responses of the two strains were compared.Results:Restriction endonuclease analyses showed that pUCluxKO-mutant vector had been successfully recombined. The Luxs-deleted status of S.mutans mutants was confirmed by PCR with primers which were specific for the genes of Luxs and Erythromycin resistance. S.mutans mutant can not induce bioluminescence, indiating the mutant had been successfully recombined. After twenty generations of culture, the constructed Chinese S.mutans mutants were confirmed to be stable. Significant difference of aciduricity was observed between S.mutans standard strain and LuxS mutant strain.The acid resistance of standard strain was stronger than that of LuxS mutant strain.The two strains both displayed the capability of acid tolerance responses. Conclusion:The S.mutans gene allelic exchange plasmid is constructed correctively and a Luxs-negative mutants of S.mutans is constructed, which can help to further study the role of Luxs in the pathogenesis of S.mutans. LuxS mutant strain is more sensitive to acid inactivation,but the capability of acid tolerance responses exist still.

抗枯萎病宝岛蕉早花突变体的筛选与鉴定

【目的】筛选鉴定抗枯萎病香蕉品种宝岛蕉的早花突变体,为进一步利用突变体株系选育适应性更广的香蕉新品种提供参考依据。【方法】利用多代组织培养繁育结合田间种植对比试验,依据生育期短、抗病性强、株高矮、株产高和遗传稳定筛选目标,从宝岛蕉早花突变后代中筛选优良株系,以宝岛蕉为对照,按照香蕉种质资源描述规范对优良突变株系的形态特征和生物学特性等进行观察鉴定,测定分析植株球茎生长点部位碳、氮营养累积以及成花相关激素吲哚乙酸(IAA)、玉米素(ZR)、赤霉素(GA)和脱落酸(ABA)含量差异,通过ISSR分子标记检测突变体及其野生型之间的遗传变异和亲缘关系。以宝岛蕉和主栽品种(桂蕉6号及巴西蕉)为对照,利用伤根浸菌法进行苗期抗病性鉴定,并在广西和海南开展区域种植试验及田间抗病性鉴定。【结果】筛选获得优良突变株系0523(简称0523),其新植蕉平均株高280.0 cm,假茎基围80.5 cm、中围60.0 cm,较宝岛蕉分别减小14.0%、5.8%和11.8%,新抽总叶片数26~29片;果实营养品质和单株产量与宝岛蕉无显著差异(P>0.05,下同)。0523球茎生长点碳氮比较宝岛蕉显著提高19.0%(P<0.05,下同),GA和IAA含量显著降低24.8%和22.6%,ABA和ZR含量与宝岛蕉相比略有增加,差异不显著。抗病性苗期鉴定结果显示0523为中抗且偏强,抗性水平与宝岛蕉相比略有提高,桂蕉6号为高感。突变株系0523与宝岛蕉的多态性比率为8.8%,遗传相似系数为0.95,二者存在差异。0523在广西和海南各试验点第1、2造蕉平均发病率分别为4.5%和3.5%,较主栽品种降低87.8%和94.1%,各试验点发病率均显著低于主栽品种。0523第1、2造蕉的生育期较宝岛蕉显著缩短,与主栽品种无显著差异。【结论】突变株系0523具有抗枯萎病、早熟、矮化、适应性广等优良性状,可用来培育大面积推广的香蕉新品种,或作为亲本育种材料应用于香蕉育种研究。

苜蓿抗寒突变体生理生化及性状指标分析

为选育在黑龙江省可越冬的理想紫花苜蓿品种。以‘龙牧806’零磁空间诱变处理后二代群体中获得的lm1609、lm1625、lm1704三个抗寒突变体为实验材料,通过测定发芽指标、越冬率、产量特性指标及根系生理生化指标,进行抗寒性能鉴定。3个突变体发芽指数与对照差异不显著;发芽势、简化活力指数lm1609显著高于lm1625、lm1704,分别为82.05%、469.82(P<0.05);3个突变体的越冬率都显著低于对照组;突变体和对照6月6日起始株高差异不显著,随着植株生长,7月6日和8月6日lm1609、lm1704显著高于lm1625和对照;突变体lm1609、lm1625、lm1704三者鲜草产量差异不显著,但显著高于对照(P<0.05);三个苜蓿突变体植株根系过氧化物酶、超氧化物歧化酶均显著高于对照,并且突变体lm1609 POD、SOD最高,分别达到304.85 OD470/(min·g)、208.41 U/g(P<0.05)。lm1609突变体具有较高的抗寒特性及产量特性,是为理想的抗寒突变体材料。

华法林耐药突变体减弱人维生素K环氧化物还原酶与华法林的结合能力

目的探讨人维生素K环氧化物还原酶(VKORC1)耐药突变体对华法林耐药的机制。方法利用与华法林结合导致VKORC1电泳迁移率变化的原理进行电泳迁移实验,并通过酶联免疫吸附实验检测VKORC1的活性和华法林抑制VKORC1的半抑制浓度(IC_(50)),评价VKORC1耐药突变体与华法林的结合能力。结果华法林与VKORC1结合后,保护其柔性跨膜螺旋1(TM1)中的一个半胱氨酸免受N-乙基马来酰亚胺(NEM)的修饰,从而增加VKORC1的电泳迁移率。增加华法林浓度可以增加具有快速迁移效应的VKORC1。与野生型VKORC1相比,华法林耐药突变体的快速迁移效应减弱或消失,表明华法林与耐药突变体的结合能力减弱;且突变体快速迁移效应越弱,其对华法林的耐药性越强。结论VKORC1耐药突变体与华法林的结合能力减弱是华法林耐药的主要原因。