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Basmati rice is known for its extra-long slender grains, exceptional kernel dimensions after cooking, high volume expansion, and strong aroma. Developing high yielding Basmati rice varieties with good cooking quality is a gigantic task. Therefore, identifying the genomic regions governing the grain and cooked kernel dimension traits is of utmost importance for its use in marker-assisted breeding. Although several QTLs governing grain dimension traits have been reported, limited attempts have been made to map QTLs for grain and cooked kernel dimension traits of Basmati rice. In the current study, a population of recombinant inbred lines (RIL) was generated from a cross of Sonasal and Pusa Basmati 1121 (PB1121). In the RIL population, there was a significant positive correlation among the length (RRL: rough rice length, MRL: milled rice length, CKL: cooked kernel length) and breadth (RRB: rough rice breadth, MRB: milled rice breadth and CKB: cooked kernel breadth) of the related traits, while there was significant negative correlation between them. QTL mapping has led to the identification of four major genomic regions governing MRL and CKL. Two QTLs co-localize with the earlier reported major gene GS3 and a QTL qGRL7.1, while the remaining two QTLs viz., qCKL3.2 (qMRL3.2) and qCKL4.1 (qMRL4.1) were novel. The QTL qCKL3.2 has been bracketed to a genomic region of 0.78 Mb between the markers RM15247 and RM15281. Annotation of this region identified 18 gene models, of which the genes predicted to encode pentatricopeptides and brassinosteroid insensitive 1-associated receptor kinase 1 precursor may be the putative candidate genes. Furthermore, we identified a novel QTL qKER2.1 governing kernel elongation ratio (KER) in Basmati rice.
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Allelic variability of the aroma gene, betaine aldehyde dehydrogenase 2 (BADH2) was studied in a random subset of indigenous aromatic rice germplasm along with a few exotic aromatic accessions. Use of functional markers of four badh2 alleles identified that the test panel possessed only two alleles, badh2-E7 and badh2-p-5'UTR. Two other alleles, badh2.2 and badh2-E4-5.2 were absent. Based on the alleles present, four functional polymorphisms (FP) were detected, namely FP1 to FP4. 188 genotypes possessed FP1 having both the badh2-p-5'UTR and badh2-E7 (71.8%) alleles. The badh2 allele with FP1 is named badh2-E7-p. 39 genotypes (14.9%) possessed only the badh2-p-5'UTR allele (FP3), while three genotypes were found to carry only the badh2-E7 allele (FP2). We also found that 32 genotypes (12.2%) did not have any of the target aroma alleles tested in this study (FP4). Interestingly, for badh2-p-5'UTR marker, the expected 198 bp amplicon for the non-aromatic allele could not be detected among any of the genotypes tested. Instead, an amplicon of 456 bp length appeared with 100% presence in the non-aromatic checks. Notwithstanding, the 456 bp allele also showed a 16% presence among the aromatic lines. This article forms the first report of this allele, named badh2-p1, among aromatic rice. Quantification of 2-Acetyl-1-Pyrroline (2AP) content and sensory evaluation among the test genotypes showed that those with FP1 are highly aromatic than the genotypes carrying other types of FPs. But, a few strongly aromatic lines showed lower 2AP content. The BADH2 characterization carried out in this study is suggestive of identifying the additional gene(s)/ allele(s) governing aroma among the Indian fragrant rice. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01181-9.
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Temperature rise predicted for the future will severely affect rice productivity because the crop is highly sensitive to heat stress at the reproductive stage. Breeding tolerant varieties is an economically viable option to combat heat stress, for which the knowledge of target genomic regions associated with the reproductive stage heat stress tolerance (RSHT) is essential. A set of 192 rice genotypes of diverse origins were evaluated under natural field conditions through staggered sowings for RSHT using two surrogate traits, spikelet fertility and grain yield, which showed significant reduction under heat stress. These genotypes were genotyped using a 50 k SNP array, and the association analysis identified 10 quantitative trait nucleotides (QTNs) for grain yield, of which one QTN (qHTGY8.1) was consistent across the different models used. Only two out of 10 MTAs coincided with the previously reported QTLs, making the remaing eight novel. A total of 22 QTNs were observed for spikelet fertility, among which qHTSF5.1 was consistently found across three models. Of the QTNs identified, seven coincided with previous reports, while the remaining QTNs were new. The genes near the QTNs were found associated with the protein-protein interaction, protein ubiquitination, stress signal transduction, and so forth, qualifying them to be putative for RSHT. An in silico expression analysis revealed the predominant expression of genes identified for spikelet fertility in reproductive organs. Further validation of the biological relevance of QTNs in conferring heat stress tolerance will enable their utilization in improving the reproductive stage heat stress tolerance in rice.
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Rapid deterioration of rice bran due to the LOX3 enzyme catalysed oxidation of PUFA is the major bottleneck for its utilization in various downstream applications. In the present study, we have identified a set of nine novel LOX3-null rice accessions carrying a deletion of C residue in the exon2 causing a frameshift mutation resulting in a truncated non-functional LOX3 protein. Our study, further manifested the predominance of C deletion based LOX3-null allele, named lox3-b, in the aromatic rice germplasm particularly in the Indian Basmati rice group. The LOX3-null genotypes exhibited significantly reduced rancidity, after six months of storage. They also showed significantly lower percentage reduction of linoleic acid (LA), higher γ-oryzanol content and lower hexanal content. A functional dCAPS marker designed based on the deletion polymorphism clearly differentiated LOX3 and lox3-b alleles, and has the potential application in marker assisted rice breeding programmes to develop cultivars with better bran storability.
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Oryza , Alelos , Genotipo , Lipooxigenasas , Oryza/genética , Fitomejoramiento , Proteínas de Plantas , Polimorfismo GenéticoRESUMEN
STUDY QUESTION: Is there an effect of male factor infertility (MFI) on either early or late morphokinetic parameters obtained during embryonic culture to blastocyst stage in a time-lapse imaging (TLI) incubator? SUMMARY ANSWER: Neither mild nor severe MFI had an impact on overall time to blastocyst or duration of individual cleavage stages in the total embryo population. WHAT IS KNOWN ALREADY: Prior studies have suggested that paternal DNA and sperm quality affect embryo morphokinetic parameters, but the impact of MFI is not fully understood. STUDY DESIGN, SIZE, DURATION: This retrospective cohort study, at a major academic fertility centre, included 536 couples (women, ≤44 years of age) undergoing IVF between September 2013 and September 2016. PARTICIPANTS/MATERIALS, SETTING, METHODS: Data from 4126 embryos cultured to the blastocyst stage in a TLI-monitored incubator were retrospectively reviewed. Embryos derived from the sperm of men with MFI were compared with those derived from patients with other infertility diagnoses. Generalized fixed and random effects models, t-test and χ2 were used as appropriate. MAIN RESULTS AND THE ROLE OF CHANCE: Couples with MFI had a higher rate of ICSI utilization and fewer usable embryos on average, and the men were older compared with couples with other diagnoses. Additionally, the women in MFI couples were younger and had higher antral follicle counts (AFCs) and higher anti-Müllerian hormone (AMH) levels compared with the other women undergoing IVF. When controlling for maternal and paternal ages, AMH and fertilization method (conventional IVF versus ICSI), neither mild nor severe MFI affected duration of individual cleavage stages or overall time to the blastocyst stage, when all or only usable embryos were examined (coefficient 0.44 hours in all embryos, P = 0.57; coefficient 0.39 hours in usable embryos, P = 0.60). Whether the sperm was surgically extracted similarly had no significant effect on embryo morphokinetic parameters. When the fertilization method was assessed independently, ICSI lengthened the overall time to blastocyst stage by 1.66 hours (P = 0.03) on average, primarily due to an increase in duration of the time from 5-cell embryo stage to early blastulation (P5SB). LIMITATIONS, REASONS FOR CAUTION: This large cohort study avoided embryo selection bias due to random assignment of embryos to the TLI incubators. However, our findings may not be generalizable to groups under-represented in our clinic population. Future studies should also evaluate the impact of male hormonal status and detailed sperm morphology, such as head versus flagellum defects, on embryo morphokinetic development. WIDER IMPLICATIONS OF THE FINDINGS: Our findings suggest that the fertilization method rather than MFI per se impacts time to early blastulation. The clinical implications of this effect on embryo development warrant further investigation. STUDY FUNDING/COMPETING INTEREST(S): There were no sources of funding for this study. There are no competing interests. TRIAL REGISTRATION NUMBER: N/A.
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Blastocisto , Técnicas de Cultivo de Embriones , Estudios de Cohortes , Femenino , Fertilización In Vitro , Humanos , Masculino , Estudios Retrospectivos , Imagen de Lapso de TiempoRESUMEN
The effect of vacuum packing and ambient storage conditions on the stability of the ß-carotene in the transgenic Golden Rice® lines was studied. The ß-carotene was quantified using RP-HPLC at bimonthly intervals for a period of six months. The ß-carotene concentration in the genotypes analyzed ranged from 7.13 to 22.81⯵g/g of endosperm. The transgene being the same in all the genotypes, variation in the ß-carotene concentration reflects on the genetic background of the rice variety and the transgene position that governed the differential accumulation of ß-carotene. It was observed that in the absence of light, oxidative degradation is higher followed by thermal degradation. Weibull model with higher R2 best explained the degradation kinetics of ß-carotene in Golden Rice® lines across all the storage conditions. The knowledge generated through this study can be utilized in devising an effective delivery system for Golden Rice® to the consumer.
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Almacenamiento de Alimentos/métodos , Oryza/metabolismo , Plantas Modificadas Genéticamente/metabolismo , beta Caroteno/metabolismo , Endospermo/genética , Endospermo/metabolismo , Embalaje de Alimentos/métodos , Cinética , Luz , Modelos Biológicos , Oryza/genética , Plantas Modificadas Genéticamente/genética , TransgenesRESUMEN
AIMS: This study aimed to evaluate the expression levels of virulence gene regulators (luxR and toxR) and virulence factors (serine protease, metalloprotease and haemolysin) in luminescent and nonluminescent isogenic Vibrio harveyi and Vibrio campbellii. METHOD AND RESULTS: Nonluminescent variants have been reported before to become dominant in cultures of luminescent vibrios when grown under static conditions in the dark. Wild-type V. harveyi BB120, V. campbellii LMG 21363, quorum sensing mutants of V. harveyi BB120 and their previously reported nonluminescent isogenic counterparts were used in this study. The expression level of the virulence genes srp serine protease, vhp metalloprotease and vhh haemolysin, the quorum sensing master regulator gene luxR and the virulence regulator gene toxR in isogenic luminescent and nonluminescent strains were quantified using reverse transcriptase real-time PCR. These experiments revealed that the nonluminescent strains produced lower levels of the quorum sensing master regulator gene luxR and the vhp metalloprotease gene (which is known to be regulated by quorum sensing). Finally, challenge tests with gnotobiotic brine shrimp (Artemia franciscana) larvae revealed that the nonluminescent strains are less virulent than their luminescent isogenic counterparts. CONCLUSION: Nonluminescent variants of V. harveyi and V. campbellii strains produce lower levels of the quorum sensing master regulator gene luxR and the vhp metalloprotease gene and are less virulent to brine shrimp than their isogenic luminescent counterparts. SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that adaptation of luminescent vibrios to specific growth conditions that result in a dominant nonluminescent phenotype is accompanied by a decreased adaptation to a host environment because of altered virulence gene regulation.
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Artemia/microbiología , Proteínas Bacterianas/metabolismo , Vibrio/patogenicidad , Factores de Virulencia/metabolismo , Animales , Artemia/crecimiento & desarrollo , Proteínas Bacterianas/genética , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Expresión Génica , Genes Reguladores , Vida Libre de Gérmenes , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Larva/microbiología , Luminiscencia , Metaloproteasas/genética , Metaloproteasas/metabolismo , Percepción de Quorum , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Serina Proteasas/genética , Serina Proteasas/metabolismo , Transactivadores/genética , Transactivadores/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vibrio/genética , Vibrio/metabolismo , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
AIMS: The study was aimed at investigating the presence of typical and atypical virulence genes in isolates belonging to the Harveyi clade (Vibrio harveyi and Vibrio campbellii). METHODS AND RESULTS: Forty-eight vibrio isolates belonging to the Harveyi clade were screened for the presence of virulence genes that are typical for these bacteria and those found in human pathogenic vibrios such as Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus and aquatic pathogenic Vibrio anguillarum. The virulence genes were amplified by PCR with specific primers, and the presence further confirmed by dot blot hybridization. The virulence genes vhh, chiA, vhpA, toxR(Vh), luxR and serine protease, typical of Harveyi clade were detected in all the isolates. The haemolysin gene hlyA and the virulence regulator gene toxR(Vc) specific to V. cholerae and the V. anguillarum-specific flagellum gene (flaC) were present in some of the isolates. Challenge tests with gnotobiotic Artemia nauplii did not show any correlation between the presence of the virulence genes and virulence of the isolates. CONCLUSION: From our results, there appears a remote possibility that vibrios belonging to the Harveyi clade might acquire virulence genes from other vibrios in the aquatic environment through horizontal gene transfer. SIGNIFICANCE AND IMPACT OF THE STUDY: Vibrios belonging to the Harveyi clade may be an important reservoir of virulence genes of other (human pathogenic) Vibrio species in the aquatic environment. The acquisition of virulence genes by horizontal transfer might increase the ability of Harveyi clade vibrios to infect aquatic organisms by increasing their virulence to a specific host by broadening their host range. The detection of such genes may forewarn the hatchery operators about a potentially virulent pathogen and thus help to develop management measures to handle the problem of vibriosis.
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Vibrio/patogenicidad , Factores de Virulencia/genética , Animales , Artemia/microbiología , Genes Bacterianos , Reacción en Cadena de la Polimerasa/métodos , Vibrio/clasificación , Vibrio/genética , Vibrio/aislamiento & purificación , Virulencia/genéticaRESUMEN
AIMS: Evaluation of protein profiling for typing Vibrio parahaemolyticus using 71 strains isolated from different seafood and comparison with other molecular typing techniques such as random amplified polymorphic DNA analysis (RAPD) and enterobacterial repetitive intergenic consensus sequence (ERIC)-PCR. METHODS AND RESULTS: Three molecular typing methods were used for the typing of 71 V. parahaemolyticus isolates from seafood. RAPD had a discriminatory index (DI) of 0.95, while ERIC-PCR showed a DI of 0.94. Though protein profiling had less discriminatory power, use of this method can be helpful in identifying new proteins which might have a role in establishment in the host or virulence of the organism. CONCLUSIONS: The use of protein profiling in combination with other established typing methods such as RAPD and ERIC-PCR generates useful information in the case of V. parahaemolyticus associated with seafood. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrates the usefulness of nucleic acid and protein-based studies in understanding the relationship between various isolates from seafood.
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Técnicas de Tipificación Bacteriana/métodos , Alimentos Marinos/microbiología , Vibrio parahaemolyticus/clasificación , Proteínas Bacterianas/metabolismo , ADN Bacteriano/genética , Electroforesis en Gel de Poliacrilamida , India , Filogenia , Reacción en Cadena de la Polimerasa/métodos , Técnica del ADN Polimorfo Amplificado Aleatorio/métodos , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/aislamiento & purificaciónRESUMEN
The successful management of BPH with minimally invasive thermal therapies requires a firm understanding of the temperature-time relationship for tissue destruction. In order to accomplish this objective, the present in vitro study assesses the cellular viability of human BPH tissue subjected to an experimental matrix of different temperature-time combinations. Hyperplastic prostate tissue was obtained from 10 radical prostatectomy specimens resected for adenocarcinoma. A portion of hyperplastic tissue from the lateral lobe of each prostate was sectioned into multiple 1 mm thick tissue strips, placed on a coverslip and thermally treated on a controlled temperature copper block with various temperatures (45-70 degrees C) for various times (1-60 min). After heat treatment, the tissue slices were cultured for 72 h and viability was assessed using two independent assays: histology and dye uptake for stromal tissue and using histology alone for the glandular tissue. The hyperplastic human prostate tissue showed a progressive histological increase in irreversible injury with increasing temperature-time severity. The dye uptake and histology results for stromal viability were similar for all temperature-time combinations. In vitro thermal injury showed 85-90% stromal destruction (raw data) of human BPH for temperature-time combinations of 45 degrees C for 60 min, 50 degrees C for 30 min, 55 degrees C for 5 min, 60 degrees C for 2 min and 70 degrees C for 1 min. Apoptosis was also identified in the control and milder treated tissues with the degree of glandular apoptosis (about 20%) more than that seen in the stromal regions (< 5%). The Arrhenius model of injury was fitted to the data for conditions leading to a 90% drop in viability (normalized to control) obtained for stromal tissue. The activation energies (E) were 40.1 and 38.4 kcal/mole for the dye uptake study and histology, respectively, and the corresponding frequency factors (A) were 1.1 x 10(24) and 7.78 x 10(22)/s. This study presents the first temperature-time versus tissue destruction relation for human BPH tissue. Moreover, it supports the concept that higher temperatures can be used for shorter durations to induce tissue injury comparable with the current clinically recommended lower temperature-longer time treatments (i.e. 45 degrees C for 60 min) for transurethral microwave thermotherapy of the prostate.
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Calor , Hipertermia Inducida/métodos , Hiperplasia Prostática/terapia , Resultado del Tratamiento , Bencimidazoles , Supervivencia Celular , Etidio/análogos & derivados , Colorantes Fluorescentes , Predicción , Humanos , Masculino , Microscopía Fluorescente , Modelos Biológicos , Próstata/patología , Hiperplasia Prostática/patología , Células del Estroma/patología , Factores de Tiempo , Técnicas de Cultivo de TejidosRESUMEN
This study examines the relationship between social support, coping resources and codependence in the wives of individuals with drug and alcohol dependence. Thirty male individuals each with drug and alcohol dependence were assessed for severity of addiction by Addiction Severity Index (ASI). Their wives were administered Social Support Scale (SSS). Coping Resources Inventory (CRI) and Codependence Assessment Questionnaire (CAQ). Of the sixty patients, forty nine were found to be codependent. On ASI, the codependent group had more impairment in the financial and the legal domains of the husbands whereas non codependent group had more impairment in the psychological domain. Codependent wives had lower coping resources and social support. On multivariate analysis, three variables were found to be significant predictors of the development of codependence in the wives. These were (i) husbands'duration of drug or alcohol abuse, (ii) total CRI score and (Hi) husbands' ASI employment score.
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A total of 405 cases of fever who were either admitted to the Hospital or attended in paediatric out patient Department or Emergency of Medical College Hospital, Calcutta between January '95 and November '95 were included in the study. Majority of cases presented with usual features of malaria like fever with chill and rigor, hepatosplenomegaly, pallor. Apart from these, complicated manifestations like shock, convulsion D.I.C and jaundice were also observed. Some unusual presentations with severe diarrhoea, dehydration and features like that of acute viral respiratory tract infection were highly confusing in terms of clinical diagnosis. P. falciparum was observed in 35.5% of cases. Overall therapeutic response to chloroquin was good, However, two patients died of cerebral Malaria. Five cases of severe malaria were caused by P. vivax however, other etiological features could not be found to attribute the severe nature of these illnesses.
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Hospitalización/estadística & datos numéricos , Malaria/epidemiología , Plasmodium falciparum/aislamiento & purificación , Plasmodium vivax/aislamiento & purificación , Animales , Antimaláricos/uso terapéutico , Niño , Preescolar , Cloroquina/uso terapéutico , Humanos , India/epidemiología , Lactante , Recién Nacido , Malaria/clasificación , Malaria/tratamiento farmacológico , Malaria/fisiopatología , Malaria Cerebral/mortalidad , Quinina/uso terapéutico , Índice de Severidad de la Enfermedad , Salud UrbanaRESUMEN
An outbreak of Dengue Haemorrhagic Fever (DHF) occurred in Calcutta between September and December, 1990. Children and young adults were the major victims. Haemorrhagic manifestations and shocks were the main features in most of the hospitalised cases. Five mouse pathogenic agents were isolated from 105 acute cases and all were identified as DEN-3. HI and CF test with 55 paired sera revealed evidence of dengue infection in 33 (60 per cent) and flavivirus group reaction including dengue in 17 (30.9 per cent). It was for the first time, that DEN-3 was considered to be the etiologic agent for DHF in Calcutta.
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Virus del Dengue/clasificación , Dengue/epidemiología , Dengue/microbiología , Brotes de Enfermedades , Vigilancia de la Población , Adolescente , Adulto , Bioensayo , Niño , Preescolar , Pruebas de Fijación del Complemento , Ensayo de Inmunoadsorción Enzimática , Femenino , Pruebas de Inhibición de Hemaglutinación , Humanos , India/epidemiología , Lactante , Masculino , Serotipificación , Población UrbanaRESUMEN
A nosocomial outbreak of neonatal septicemia due to K. pneumoniae occurred in nursery during June-July, 1991. Klebsiella pneumoniae (Klebocin type 314) was recovered from blood of 33 (70.2%) of 47 neonates with septicemia. Multiple drug resistance was observed in all the cases. The same strain of K. pneumoniae was recovered from the neonates and environment of nursery and labour room as well. The outbreak was attributable to environmental dissemination.
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Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/aislamiento & purificación , Sepsis/microbiología , Infección Hospitalaria , Brotes de Enfermedades , Farmacorresistencia Microbiana , Humanos , Recién Nacido , Klebsiella pneumoniae/efectos de los fármacos , Salas Cuna en HospitalRESUMEN
Rhizobium sp. isolated from the root nodules of Erythrina indica Lamk., a leguminous tree, produced large amounts of indoleacetic acid (IAA) from tryptophan. L-Tryptophan (2 mg/ mL) was preferred to DL-tryptophan for IAA synthesis. Attempts were made to optimize the cultural requirements for an accelerated IAA production. An enrichment of the carbon-free incubation medium with maltose (1 %), MnSO4 x H2O (5 microg/mL), and Triton X-100 (100 ppm) promoted the synthesized IAA content of the medium by 480 %.