Pyramiding and evaluation of segregating lines containing lectin and protease inhibitor genes for aphid resistance in Brassica juncea-Supplementary Data

Aphids are one of the most devastating pests, affecting the potential yield and quality ofBrassica juncea. In the current study, we have attempted to pyramid two transgenic lines containing chickpea lectin (CHPL, P1) and urdbean protease inhibitor (UPI, P2) in each under the phloem specific rolC promoter, through conventional breeding approach. In the derived F2 population, both lectin and protease inhibitor genes were segregating in a 9:3:3:1 ratio (p-value: 0.81), indicative of a single copy of the transgenes in the parents. Furthermore, the parental, as well as pyramided progenies were evaluated for their potential resistance to aphids in terms of mortality and natality. The lines containing both the transgenes were found to be superior over single gene transgenics as a higher mortality rate (96%) was found in F2on the 9th day as compared to single gene transgenics (86% and 80% in P1 and P2 respectively). A significant decrease in the number of nymphs was observed in P1 and P2 but most in F2 plants as almost 43, 32.08, and 107.5 times decrease in the number of nymphs was found in P1, P2, and F2 individuals over control. Expression profiling was done to see if there was any impact of gene pyramiding on the expression pattern of both transgenes before and after aphid treatment, and no significant changes were observed, indicating constitutive expression of transgenes in pyramided lines also. In conclusion, pyramided lines were found to be promising and were superior for aphid resistance.

Pyramiding and evaluation of segregating lines containing lectin and protease inhibitor genes for aphid resistance in Brassica juncea

Aphids are one of the most devastating pests, affecting the potential yield and quality ofBrassica juncea. In the current study, we have attempted to pyramid two transgenic lines containing chickpea lectin (CHPL, P1) and urdbean protease inhibitor (UPI, P2) in each under the phloem specific rolC promoter, through conventional breeding approach. In the derived F2 population, both lectin and protease inhibitor genes were segregating in a 9:3:3:1 ratio (p-value: 0.81), indicative of a single copy of the transgenes in the parents. Furthermore, the parental, as well as pyramided progenies were evaluated for their potential resistance to aphids in terms of mortality and natality. The lines containing both the transgenes were found to be superior over single gene transgenics as a higher mortality rate (96%) was found in F2on the 9th day as compared to single gene transgenics (86% and 80% in P1 and P2 respectively). A significant decrease in the number of nymphs was observed in P1 and P2 but most in F2 plants as almost 43, 32.08, and 107.5 times decrease in the number of nymphs was found in P1, P2, and F2 individuals over control. Expression profiling was done to see if there was any impact of gene pyramiding on the expression pattern of both transgenes before and after aphid treatment, and no significant changes were observed, indicating constitutive expression of transgenes in pyramided lines also. In conclusion, pyramided lines were found to be promising and were superior for aphid resistance.

Biochemical characterization for determination of genetic distances among different indigenous chickpea (Cicer arietinum L. ) varieties of North-West Pakistan / Caracterização bioquímica para determinação de distâncias genéticas entre diferentes variedades indígenas de grão-de-bico (Cicer arietinum L. ) do noroeste do Paquistão

Abstract Genetic distances among different chickpea varieties and evaluation of their free amino acid profiles were determined on the basis of Sodium dodecyle sulphate polyacrylamide gels electrophoresis (SDS-PAGE). Total soluble proteins were resolved on 10% SDS Polyacrylamide gel. Low variability in tested varieties was observed. Dendogram based on electrophoretic data clustered the genotypes into 2 groups. The results showed that the average protein content of all the varieties was 26.01% within the range 22.8% for Thal-2006 to 34.06% Sheenghar-2000 of dry seed weight. On the basis of total protein content Bittal-98, Dasht and Sheen Ghar-2000, Karak-3 and CM-98, Paidar -91 and Fakhr-e-Thal, C-44, Balaksar and KK-1showed similar concentrations for protein contents among each other but showed variation from the rest of the varieties. Different proteins were separated on the basis of changes in their molecular weights by means of Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Dasht, CM-98, and Sheen Ghar showed 100% similarity. Balaksar and Fakhr-e- Thal, KK-2 and Chattan and KC-98, KK-1 and Lawaghar were 100% similar among each other but showed variation from the rest of the accessions. The overall dendrogram showed high and low level of variation among the accessions. The concentration of free amino acids varied among the 16 chickpea varieties. A significant difference of both essential and non-essential amino acids was found among the chickpea cultivars. The total concentration of essential amino acid was recorded 40.81 g/100 g protein while non-essential was recorded 59.18343 g/100 g protein in the given cultivars. The highest concentration of essential amino acids was found in C-44 followed by KK-2, KK-1 and Fakhr E Tal while the lowest concentration was recorded in Cm-98, Paidar-91 and Sheen Ghar-2000 respectively. Cultivars TAL-2006, Chattan and Karak-3 showed maximum concentration of both essential and endogenous amino acids. In conclusion; for broadening the genetic pools in breeding programs or to search for exotic characters, for instance new disease resistance alleles, accession with low similarity coefficients (Lawaghar and Battal-98) may be utilized. Furthermore the information acquired from this study could be used to device a proficient breeding approach intended at improving nutritional as well as broadening the genetic base of this essential food crop of Pakistan.

Resumo As distâncias genéticas entre as diferentes variedades de grão-de-bico e a avaliação de seus perfis de aminoácidos livres foram determinadas com base na eletroforese em gel de poliacrilamida com dodecil sulfato de sódio (SDS-PAGE). As proteínas solúveis totais foram resolvidas em SDS-PAGE a 10%. Foi observada baixa variabilidade nas variedades testadas. O dendrograma fundamentado em dados eletroforéticos agrupou os genótipos em dois grupos. Os resultados mostraram que o teor médio de proteínas de todas as variedades foi de 26,01%, na faixa de 22,8% para Thal-2006 a 34,06% para Sheenghar-2000 do peso de sementes secas. Com base no conteúdo total de proteínas, Bittal-98, Dasht, Sheen Ghar-2000, Karak-3, CM-98, Paidar-91, Fakhr-e-Thal, C-44, Balaksar e KK-1 apresentaram concentrações semelhantes para o conteúdo de proteínas entre si, mas tiveram variação quanto ao restante das variedades. Diferentes proteínas foram separadas com base nas alterações de seus pesos moleculares por meio de eletroforese em gel de poliacrilamida com dodecil sulfato de sódio (SDS-PAGE). Dasht, CM-98 e Sheen Ghar mostraram 100% de similaridade. Balaksar, Fakhr-e-Thal, KK-2, Chattan e KC-98, KK-1 e Lawaghar foram 100% semelhantes entre si, mas apresentaram variação em relação ao restante dos acessos. O dendrograma geral mostrou alto e baixo nível de variação entre os acessos. A concentração de aminoácidos livres variou entre as 16 variedades de grão-de-bico. Foi encontrada uma diferença significativa entre os aminoácidos essenciais e não essenciais nas cultivares de grão-de-bico. A concentração total de aminoácidos essenciais foi registrada em 40,81 g / 100 g de proteína, enquanto a não essencial foi registrada em 59,18343 g / 100 g de proteína nas cultivares. A maior concentração de aminoácidos essenciais foi encontrada em C-44, seguida de KK-2, KK-1 e Fakhr-e-Thal, enquanto a menor concentração foi registrada em CM-98, Paidar-91 e Sheen Ghar-2000. As cultivares TAL-2006, Chattan e Karak-3 apresentaram concentração máxima de aminoácidos essenciais e endógenos. Em conclusão, para ampliar os pools genéticos em programas de melhoramento ou procurar caracteres exóticos, por exemplo, novos alelos de resistência a doenças, pode ser utilizada a adesão com baixos coeficientes de similaridade (Lawaghar e Battal-98). Além disso, as informações adquiridas neste estudo poderiam ser usadas para criar uma abordagem de criação eficiente, com o objetivo de melhorar a nutrição e ampliar a base genética dessa cultura alimentar essencial do Paquistão.

Whole chickpea flour as an ingredient for improving the nutritional quality of sandwich bread: Effects on sensory acceptance, texture profile, and technological properties

ABSTRACT The inclusion of pulse flours with higher dietary fiber content in bakery products, such as chickpea (Cicer arietinum L) flour, has been exploited by the food industry due to its nutritional and sensory characteristics. This work aimed to develop a sandwich bread with a partial substitution (7.5%, 15%, and 30%) of refined wheat flour by whole chickpea flour (WCF) and also to evaluate the effects on sensory acceptance, physicochemical parameters, and texture profile during the shelf life. Four methods for obtaining WCF were assessed and the best condition was used to produce the sandwich bread. Sensory evaluation was carried out with 65 consumers using a 9-point hedonic scale and a purchase intention scale. Firmness, elasticity, cohesiveness, chewability, and fracturability was evaluated by the Texture Profile Analysis (TPA). The specific volume, total fungi count, pH, titratable acidity, and moisture content were also determined. The nutritional composition was estimated by mass balance. Data demonstrated that there was no difference (p>0.05) in sensory evaluation of the products elaborated with different concentrations of WCF, and all samples presented good acceptance. The addition of WCF did not cause negative effects on the texture and volume characteristics of the bread. The sample with 30% WCF had adequate technological characteristics, resulting in a product with higher nutritional value, especially dietary fiber, and was well accepted by the consumers. This formulation can be used by the food industry and also in domestic preparations, contributing to the diversification and nutritional enrichment of sandwich bread.

RESUMEN La inclusión de harinas de granos no convencionales con mayor contenido de fibra dietética en productos de panadería, como la harina integral de garbanzo (HIG) (Cicer arietinum L), es utilizada por la industria alimentaria debido a sus características nutricionales y sensoriales. Este trabajo tuvo como objetivo desarrollar un pan para sándwich (PPS) con sustitución parcial (7.5%, 15% y 30%) de harina de trigo refinada por HIG así como evaluar la aceptación sensorial, parámetros fisicoquímicos y el perfil de textura durante su vida útil. Fueron evaluados cuatro métodos para la obtención de HIG, utilizándose la mejor condición para producir el PPS. Sesenta y cinco consumidores realizaron la evaluación de aceptación sensorial e intención de compra con una escala hedónica de 9 puntos. La firmeza, la elasticidad, la cohesión, la masticabilidad y la fracturabilidad se evaluaron mediante el análisis de perfil de textura (TPA). También se determinó el volumen específico, el recuento total de hongos, el pH, la acidez titulable y el contenido de humedad. La composición nutricional se estimó por balance de masa. Todas las muestras presentaron buena aceptabilidad sin diferencias estadística. La adición de HIG no causó efectos negativos en la textura ni el volumen del PPS. La muestra con 30% de HIG tubo características tecnológicas adecuadas, resultando en un producto con mayor valor nutricional, especialmente por la fibra dietética. Esta formulación podría ser utilizada por la industria alimentaria y también en preparaciones domésticas, contribuyendo a la diversificación y al enriquecimiento nutricional del pan para sándwich.

Biochemical and cytogenetic effects of Imazethapyr on Cicer arietinum L.

Weeds are a serious obstacle to the production of chickpea. Imazethapyr (IM) herbicide is used to controlweeds in most of the pulses, including chickpea. Mitotic abnormalities, chromosomal behavior, and proteincontent in chickpea (Cicer arietinum L.), due to IM treatment, were studied. The chickpea seeds (varietyJG-11) were germinated in sterilized Petri dishes, 9 cm in diameter, on Whatman filter paper moistened with10 ml of either Hoagland nutrient solution (control) or five concentrations of IM (0.5, 1, 2.5, 5, and 10 ppm).From the cytologic point of view, observations demonstrated that the mitotic frequency in root meristematiccells diminished, and that abnormality frequency increased parallel to the increase in concentrations of IM.The herbicide was highly mito-inhibitory and induced chromosomal irregularities, such as stickiness, lagging,scattering, and chromosome bridges. The endosperm and root-shoot axis’ protein content decreased withincreasing of the herbicide concentration in all the treatments. It can be argued that IM produces undesirableside effects during mitosis in chickpea’s somatic cells and biochemical parameters

Effect of Seed Hydropriming on Seedling Emergence and Growth of Chickpea (Cicer arietinum L.) under Adverse Climatic Condition of Drought and Salinity

Aim: Assessment of influence of seed hydropriming in seed and seedling vigour enhancement for stress tolerance in chickpea which is important for successful crop production under erratic climate change causing drought and salinity stress severely affecting the seedling emergence and establishment especially in arid and semi arid regions of the world. Study Design: A three factorial randomised block design was used in the study, involving seed hydropriming treatment, chickpea varieties and growth conditions. Place and Duration of the Study: The experiment was conducted in Department of Seed Science and Technology, University of Agricultural Sciences, Dharwad, India, during the year 2018. Methodology: Six months old chickpea seeds of variety JG-11 and Annigeri-1 were hydroprimed for 12 hours at 25°C in dark condition. The primed and unprimed control seeds are sown under normal, drought and salinity conditions imposed in pots under controlled conditions. The data recorded were analysed for Analysis of Variance (ANNOVA). Results: The result of the experiment revealed that, upon seed hydropriming an average of 19 per cent increase in seedling emergence and 53.3 per cent increase in seedling vigour index was observed as compared to unprimed control seeds irrespective of variety and the abiotic stress conditions.

Ecofriendly Management of Seedling Diseases of Chickpea (Cicer arietinum)

The experiment was carried out in the field of Plant Pathology Department, Bangladesh Agricultural University, Mymensingh, Bangladesh to determine the effect of BARI-biofertilizerand Integrated Pest Management (IPM) biopesticide for controlling foot and root rot diseases of chickpea. It was observed that both BARI-Biofertilizer and IPM biopesticide resulted significantly lower disease incidence of seedlings of the test pulse over the control. Soil treatment with BARI-Biofertilizer resulted the lowest disease incidence of chickpea var. Hyprosola, Binasola-2, Binasola-3 and Binasola-4 at 20 DAS (Days after sowing) that displayed reduction of disease incidence up to 83.77%, 54.48%, 70.76% and 71.45% respectively over control. While at 28 DAS, showedup to 82.82%, 71.92%, 84.72% and 68.39%, respectively, reduction of disease incidence over control. At 35 DAS, exhibited up to 79.91%, 73.18%, 81.32% and 73.44%, respectively, reduction of disease incidence over control. BARI-biofertilizer and IPM biopesticide increased fresh weight of plant, number of nodules per plant and fresh weight of nodules per plant.

Effect of foliar fertilization with zinc and manganese sulfate on yield, dry matter accumulation, and zinc and manganese contents in leaf and seed of chickpea (Cicer arietinum)

To investigate the effects of zinc (Zn) and manganese (Mn) foliar fertilization on yield, dry matter accumulation,Zn and Mn concentrations in leaf and seed of chickpea cultivars, a field experiment was conducted in 2014. Theexperimental design was a split plot in randomized complete block with three replications. The phenological stagesrecorded were as follows: Emergence, flowering, 50% flowering, and harvest maturity. At harvest, economic yieldand its components, biological yield, harvest index, stem, leaf, pod, seed, and total dry weight were measured. TheZn and Mn contents were determined by atomic absorption spectrometry. The results indicated that the spraying ofZn and Mn elements did not have a significant effect on the time from emergence to 50% flowering and the timefrom emergence to maturation. Spraying treatments had a significant effect on dry weight of stems, leaves, pods,seeds, and total plant. In all three cultivars, Zn spraying had the greatest effect on the plant height, number of pods perplant, number of seeds per plant, 100-seed weight, grain yield, biological yield, and dry weight of leaves, seeds, andtotal plant, while Mn spraying had the greatest effect on the increase in stem and pod dry weight and protein content.

Comparative physiological and leaf proteome analysis between drought-tolerant chickpea Cicer reticulatumand drought-sensitive chickpea C. arietinum

Comparative physiological and proteomic analysis were performed to understand the stress responses of two chickpeaspecies (C. reticulatum and C. arietinum) against drought. Our study revealed that drought stress reduced root length, leafwater content, and enhanced free proline content in both species. Effect of drought stress appeared to be greater in C.arietinum compared to C. reticulatum. A total of 24 differently expressed proteins were identified by using MALDI-TOF/TOF-MS/MS in response to drought. The proteins involved in photosynthesis and energy mechanisms were up-regulated inC. reticulatum and down-regulated in C. arietinum under drought. Our results suggest that the photosynthesis capacity of C.reticulatum is greater than that of C. arietinum under drought stress. Abundance of proline and sucrose biosynthesis relatedproteins, glutamine synthetase and cyctosolic fructose-bisphosphate aldolase, respectively, also increased in C. reticulatumunder drought stress. The findings of this proteome analysis will help in understanding the mechanism of drought resistancein chickpea and may be also helpful in developing drought-resistant transgenic plants.

Weather based prediction model for beet armyworm (Spodoptera exigua Hubner) in chickpea

Aim: The study aimed to develop and validate weather based prediction model for beet armyworm (Spodoptera exigua) population in chickpea through adult catches in pheromone traps. Methodology: The data on adult trap catches of S. exigua were recorded daily and weekly means were computed. Log transformed trap catches data were used for correlation with weather parameters of current week, 1-lag, 2-lag and 3-lag weeks. Thereafter, multiple-linear regression analysis was done and a model was developed. The prediction model of S. exigua was validated with the appropriate statistical tools. Results: Peak incidence of S. exigua was recorded during 45th standard meteorological week (SMW) with 15.6 moths per trap per week. Amongst current, 1-lag, 2-lag and 3-lag week weather parameters, the male moth population had significant positive correlation with maximum temperature (Tmax) and minimum temperature (Tmin), and negative correlation with morning relative humidity (RH1) of 2-lag week. The sunshine hours/day (SSH) of current week had a significant negative association with S. exigua male moth catches, while the soil temperature (ST) of 2-lag week had highest positive correlation with trap catches. Regression equation was computed by regressing male moth catches of S. exigua against weather data of weeks with highest correlation coefficient. Interpretation: Often, pest-weather models are developed based on current week weather factors. However, it has been witnessed in this study that weather of preceding weeks (up to 3-lag) may also influence the pest population, and thus it needs to be considered for proper understanding of pest dynamics

Assessment of flatulence causing agents in Chickpea (Cicer arietinum L.) and their possible removal

Abstract Flatulence and fullness of stomach is one of the most common problem associated with chickpea primary due to presence of some oligosaccharides and phenols. In this investigation Desi and Kabuli varieties were compared for these oligosaccharides and phenolic compounds. Furthermore, the effect of different processing and cooking methods such as soaking, cooking and germination in the reduction of these antiphysiological factors were are also studies. Maximum tannic acid (0.90 ± 0.20%) was observed in Parbat and C-44 while minimum (0.60 ± 0.04%) in Karak-2. Stachyose contents ranged between 1.10 ± 0.05 (Karak-3) to 1.42 ± 0.02% (Parbat) while raffinose was 0.63 ± 0.05(Karak-3) to 0.81 ± 0.02% (Dasht). The highest tannic acid content was reduced up to 50% in C-44 by cooking of 72 hours germinated seeds. Stachyose and raffinose contents were completely removed after 72 hours germination. Present studies revealed that cooking after germination is the most effective method to reduce the anti-nutritional factors of chickpea. Individually, soaking and cooking also contributed to the loss of the same factors but to a lesser extent.

Biochemical characterization for determination of genetic distances among different indigenous chickpea (Cicer arietinum L.) varieties of North-West Pakistan

Abstract Genetic distances among different chickpea varieties and evaluation of their free amino acid profiles were determined on the basis of Sodium dodecyle sulphate polyacrylamide gels electrophoresis (SDS-PAGE). Total soluble proteins were resolved on 10% SDS Polyacrylamide gel. Low variability in tested varieties was observed. Dendogram based on electrophoretic data clustered the genotypes into 2 groups. The results showed that the average protein content of all the varieties was 26.01% within the range 22.8% for Thal-2006 to 34.06% Sheenghar-2000 of dry seed weight. On the basis of total protein content Bittal-98, Dasht and Sheen Ghar-2000, Karak-3 and CM-98, Paidar -91 and Fakhr-e-Thal, C-44, Balaksar and KK-1showed similar concentrations for protein contents among each other but showed variation from the rest of the varieties. Different proteins were separated on the basis of changes in their molecular weights by means of Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Dasht, CM-98, and Sheen Ghar showed 100% similarity. Balaksar and Fakhr-e- Thal, KK-2 and Chattan and KC-98, KK-1 and Lawaghar were 100% similar among each other but showed variation from the rest of the accessions. The overall dendrogram showed high and low level of variation among the accessions. The concentration of free amino acids varied among the 16 chickpea varieties. A significant difference of both essential and non-essential amino acids was found among the chickpea cultivars. The total concentration of essential amino acid was recorded 40.81 g/100 g protein while non-essential was recorded 59.18343 g/100 g protein in the given cultivars. The highest concentration of essential amino acids was found in C-44 followed by KK-2, KK-1 and Fakhr E Tal while the lowest concentration was recorded in Cm-98, Paidar-91 and Sheen Ghar-2000 respectively. Cultivars TAL-2006, Chattan and Karak-3 showed maximum concentration of both essential and endogenous amino acids. In conclusion; for broadening the genetic pools in breeding programs or to search for exotic characters, for instance new disease resistance alleles, accession with low similarity coefficients (Lawaghar and Battal-98) may be utilized. Furthermore the information acquired from this study could be used to device a proficient breeding approach intended at improving nutritional as well as broadening the genetic base of this essential food crop of Pakistan.

Resumo As distâncias genéticas entre as diferentes variedades de grão-de-bico e a avaliação de seus perfis de aminoácidos livres foram determinadas com base na eletroforese em gel de poliacrilamida com dodecil sulfato de sódio (SDS-PAGE). As proteínas solúveis totais foram resolvidas em SDS-PAGE a 10%. Foi observada baixa variabilidade nas variedades testadas. O dendrograma fundamentado em dados eletroforéticos agrupou os genótipos em dois grupos. Os resultados mostraram que o teor médio de proteínas de todas as variedades foi de 26,01%, na faixa de 22,8% para Thal-2006 a 34,06% para Sheenghar-2000 do peso de sementes secas. Com base no conteúdo total de proteínas, Bittal-98, Dasht, Sheen Ghar-2000, Karak-3, CM-98, Paidar-91, Fakhr-e-Thal, C-44, Balaksar e KK-1 apresentaram concentrações semelhantes para o conteúdo de proteínas entre si, mas tiveram variação quanto ao restante das variedades. Diferentes proteínas foram separadas com base nas alterações de seus pesos moleculares por meio de eletroforese em gel de poliacrilamida com dodecil sulfato de sódio (SDS-PAGE). Dasht, CM-98 e Sheen Ghar mostraram 100% de similaridade. Balaksar, Fakhr-e-Thal, KK-2, Chattan e KC-98, KK-1 e Lawaghar foram 100% semelhantes entre si, mas apresentaram variação em relação ao restante dos acessos. O dendrograma geral mostrou alto e baixo nível de variação entre os acessos. A concentração de aminoácidos livres variou entre as 16 variedades de grão-de-bico. Foi encontrada uma diferença significativa entre os aminoácidos essenciais e não essenciais nas cultivares de grão-de-bico. A concentração total de aminoácidos essenciais foi registrada em 40,81 g / 100 g de proteína, enquanto a não essencial foi registrada em 59,18343 g / 100 g de proteína nas cultivares. A maior concentração de aminoácidos essenciais foi encontrada em C-44, seguida de KK-2, KK-1 e Fakhr-e-Thal, enquanto a menor concentração foi registrada em CM-98, Paidar-91 e Sheen Ghar-2000. As cultivares TAL-2006, Chattan e Karak-3 apresentaram concentração máxima de aminoácidos essenciais e endógenos. Em conclusão, para ampliar os pools genéticos em programas de melhoramento ou procurar caracteres exóticos, por exemplo, novos alelos de resistência a doenças, pode ser utilizada a adesão com baixos coeficientes de similaridade (Lawaghar e Battal-98). Além disso, as informações adquiridas neste estudo poderiam ser usadas para criar uma abordagem de criação eficiente, com o objetivo de melhorar a nutrição e ampliar a base genética dessa cultura alimentar essencial do Paquistão.

Establishment and optimization of systems for protoplasts isolation of soybean and chickpea that used in subcellular location / 生物工程学报

Young leaves of Kabuli chickpea as well as soybean Xiangdou No.3, which are the current plants that studied in our laboratory were selected as materials. Effects on protoplasts yield and survival rate of different enzyme combination, concentration of D-Mannitol in enzyme combinations, pH of enzyme combinations and enzymolysis time are detected. The results showed that, the best condition for Xiangdou No.3 leaf protoplasts isolation is to rotate the cut materials for 6 hours in enyzme solution under temperature of 27 ℃ and rotate speed of 45 r/min for 6 h. Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%) in combination with Pectolyase Y-23 (0.4%) dissolving in CPW solution with MES (0.1%) and Mannitol (10%), pH 6.0 was found best for protoplasts isolation of Xiangdou No.3 leaves.The best condition for protoplasts isolation of Kabuli chickpea is to put the cut materials into enzymatic hydrolysate enzymolyse for 7 to 8 hours under temperature of 27 ℃ and rotate speed of 45 r/min on water bath shaker, the optimum combination of enzyme consists of Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%), MES (0.1%) and Mannitol (10%) dissolved in CPW solution with pH 4.8. The protoplasts prepared with the methods above are used in subcellular location and the effects show well.

Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea

Abstract The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20 °C to 40 °C, pH range of 7–11 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40) but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40), hydrocyanic acid (except VAI-7 and VAI-40), indole acetic acid and β-1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and β-1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea.

Antihyperlipidemic activity of chickpea sprouts supplementation in ovariectomy‑induced dyslipidemia in rats.

Background: Phytoestrogens are increasingly becoming popular as alternatives for hormone replacement therapy in postmenopausal condition. Objective: In this study, the antihyperlipidemic effect of chickpea (Cicer arientum) sprouts was evaluated in ovariectomy‑induced dyslipidemia in rat model in comparison with standard antihyperlipidemic agent atorvastatin. Materials and Methods: A total of 24 female adult Wistar rats were divided into four groups that is, Group I ‑ Control; Group II ‑ Ovariectomized (OVX) rats; Group III ‑ OVX + germinated chickpea sprouts (20% in diet) and Group IV OVX + atorvastatin (1.2 mg/kg b.wt, p.o.). Body and organ weights, serum, and liver lipid profile were assessed at the end of 8 weeks. Results: The results indicated that ovariectomy significantly (P < 0.05) increased total cholesterol, nonhigh‑density lipoprotein cholesterol and triglycerides (TGs) in serum and liver. The total lipid and phospholipid content in liver were also significantly (P < 0.05) increased. The weights of uterus and heart were significantly (P < 0.05) decreased. Dietary supplementation with germinated chickpea normalized the lipid profile in serum and liver. Further, high‑density lipoprotein (HDL) cholesterol, body weight, uterine, heart, and spleen weights were significantly (P < 0.05) increased. Atorvastatin administration showed similarly normalized lipid profile, but showed no improvement on decreased uterus and heart weights. Histopathological examination revealed fatty changes in liver, uterine atrophy, and subintimal fat accumulation in aorta in OVX group. The changes were mild in chickpea group with no improvement in statin group. Conclusions: Germinated seeds of chickpea showed significant antihyperlipidemic activity, which was comparable to atorvastatin. Further, germinated chickpea improved organ weights and helped in the reversal of histopathological changes suggesting its usefulness in postmenopausal condition.

Study on Synergistic Hypoglycemic ActivityIn Vitroof Three Kinds of Isoflavone in Chickpea and Its Derivatives of Ethoxylation / 世界科学技术-中医药现代化

Through the ethoxylation structure modification of three isoflavones which were genistein, biochanin A and formononetin in chickpea, the hypoglycemic activity and the synergistic hypoglycemic activity were studied. Ethoxylation structure modification was given on three kinds of isoflavones. The hypoglycemic activity of three kinds of isoflavones and their derivatives were studied. The synergistic hypoglycemic activity of the compounds was also studied. The insulin-resistance HepG2 cell was selected as the model of hypoglycemic activity screening. The results showed that four ethoxylation products were synthesized. And the hypoglycemic activity of genistein was better than biochanin A and formononetin with significant difference (P 0.05). Effects of the compound b and compound c were not as good as the compound a and compound d. There was statistical difference (P 0.05). It was concluded that the combinations of compounds played synergistic effects with better hypoglycemic effect. It provided a basis for developing hypoglycemic agents with the intellectual property rights.

DPPH radical scavenging activity and contents of H2O2, malondialdehyde and proline in determining salinity tolerance in chickpea seedlings.

The involvement of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and contents of H2O2, malondialdehyde (MDA) and proline was investigated in determining salinity tolerance among seedlings of thirty chickpea (Cicer arietinum L.) genotypes having different pedigrees. Chickpea genotypes, including cultivars and advanced lines were grown for 7 days under control and salt stress (50 mM NaCl) conditions. The genotypes showed differential response to salt stress in terms of growth, DPPH radical scavenging activity and contents of H2O2, MDA and proline in seedlings. On the basis of seedling growth, the genotypes having better performance under stress conditions had reduced levels of H2O2 and MDA contents, but increased levels of proline and DPPH radical scavenging activity. Stress tolerance index for these parameters was also determined. Agglomerative hierarchal clustering by Pearson correlation coefficient grouped the genotypes into two major clusters — MC I and MC II. MC II and A1-1 sub-cluster of MC-I comprised mainly of genotypes that showed higher stress resistance levels for the respective parameters in comparison to genotypes in other sub-clusters. Thus, it is possible to identify salt-tolerant genotypes on the basis of above parameters without a field trial.

Androgenesis in chickpea: Anther culture and expressed sequence tags derived annotation.

Double haploid technique is not routinely used in legume breeding programs, though recent publications report haploid plants via anther culture in chickpea (Cicer arietinum L.). The focus of this study was to develop an efficient and reproducible protocol for the production of double haploids with the application of multiple stress pre-treatments such as centrifugation and osmotic shock for genotypes of interest in chickpea for their direct use in breeding programs. Four genotypes, ICC 4958, WR315, ICCV 95423 and Arearti were tested for anther culture experiments. The yield was shown to be consistent with 3-5 nucleate microspores and 2-7 celled structures with no further growth. To gain a further insight into the molecular mechanism underlying the switch from microsporogenesis to androgenesis, bioinformatics tools were employed. The challenges on the roles of such genes were reviewed while an attempt was made to find putative candidates for androgenesis using Expressed Sequenced Tags (EST) and interolog based protein interaction analyses.

Pathogenicity Tests and Evaluation of Efficacy of Fungicides Against Rhizoctonia Bataticola, the Causal Agent of Dry Root Rot of Chickpea.

The pathogen was identified based on its mycelial and sclerotial characters and pathogenicity test was proved by soil inoculation method. Efficacy of two non systemic fungicides (copper oxychloride and captan), two systemic fungicides (hexaconazole and tebuconazole) and one antifungal antibiotic validamycin each at different concentrations were tested against Rhizoctonia bataticola, incitant of dry root rot of chickpea under in vitro conditions. The fungicides copper oxychloride, captan, hexaconazole and tebuconazole were found to be highly effective (100%) in inhibiting the mycelial growth of the highly virulent pathogen at all the concentrations tested.

Starches from non - conventional sources to improve the technological characteristics of pound cake / Amido de fontes não - convencionais para melhorar as características tecnológicas de bolo inglês

This study evaluated and compared the effect of the utilization of five different non-conventional starches (chickpea, common bean, Peruvian carrot, sweet potato and white bean) and four different commercial starches (cassava, corn, potato and rice) in pound cake. Common bean starch, followed by Peruvian carrot starch were the non-conventional starch sources that showed tendency to improve the technological quality of pound cake, mainly in relation to corn starch, the most common commercial source. With these sources, the batters presented lower specific gravity and the cakes presented higher specific volume, lighter color, lower crumb moisture reduction during the storage period, and better texture attributes during all the cake shelf-life. Moreover, common bean starch provided higher scores in the cake sensory evaluation; especially for grain and texture attributes (moisture, tenderness and softness). Chickpea and white bean starches were more similar to corn starch in pound cake application.

Este estudo avaliou e comparou o efeito da utilização de cinco amidos de fontes não convencionais (grão de bico, feijão carioca, mandioquinha, batata doce e feijão branco) e de quatro diferentes amidos de fontes comerciais (mandioca, milho, batata e arroz) em bolo inglês. Amido de feijão carioca, seguido pelo amido de mandioquinha, foram os amidos de fontes não convencionais que mostraram tendência a melhorar a qualidade tecnológica do bolo inglês, principalmente em comparação com o amido de milho, o qual é a fonte comercial mais utilizada. Com o amido dessas fontes, os batidos apresentaram menores valores de gravidade específica e os bolos maiores valores de volume específico, cor mais clara, menor redução da umidade durante o período de estocagem e melhores valores nos atributos de textura durante a vida de prateleira. Além disso, amido de feijão carioca promoveu maiores escores na avalição sensorial; especialmente para os atributos do miolo e textura (umidade, suavidade e maciez). Amidos de grão de bico e feijão branco foram muito similares ao amido de milho em aplicação em bolo inglês.