Application of biotechnology in breeding lentil for resistance to biotic and abiotic stress

Lentil is a self-pollinating diploid (2n = 14 chromosomes) annual cool season legume crop that is produced throughout the world and is highly valued as a high protein food. Several abiotic stresses are important to lentil yields world wide and include drought, heat, salt susceptibility and iron deficiency. The biotic stresses are numerous and include: susceptibility to Ascochyta blight, caused by Ascochyta lentis; Anthracnose, caused by Colletotrichum truncatum; Fusarium wilt, caused by Fusarium oxysporum; Sclerotinia white mold, caused by Sclerotinia sclerotiorum; rust, caused by Uromyces fabae; and numerous aphid transmitted viruses. Lentil is also highly susceptible to several species of Orabanche prevalent in the Mediterranean region, for which there does not appear to be much resistance in the germplasm. Plant breeders and geneticists have addressed these stresses by identifying resistant/tolerant germplasm, determining the genetics involved and the genetic map positions of the resistant genes. To this end progress has been made in mapping the lentil genome and several genetic maps are available that eventually will lead to the development of a consensus map for lentil. Marker density has been limited in the published genetic maps and there is a distinct lack of co-dominant markers that would facilitate comparisons of the available genetic maps and efficient identification of markers closely linked to genes of interest. Molecular breeding of lentil for disease resistance genes using marker assisted selection, particularly for resistance to Ascochyta blight and Anthracnose, is underway in Australia and Canada and promising results have been obtained. Comparative genomics and synteny analyses with closely related legumes promises to further advance the knowledge of the lentil genome and provide lentil breeders with additional genes and selectable markers for use in marker assisted selection. Genomic tools such as macro and micro arrays, reverse genetics and genetic transformation are emerging technologies that may eventually be available for use in lentil crop improvement.

Association of Dominant and Recessive Genes Confers Anthracnose Resistance in Stem and Leaves of Common Bean

Different genes might be involved in Colletotrichum lindemuthianum resistance in leaves and stem of common bean. This work aimed to study the genetic mechanisms of the resistance in the leaf and stem in segregating populations from backcrosses involving resistant cultivar AN 910408 and susceptible cultivar Ruda inoculated with spore suspensions of C. lindemuthianum race 83. Our results indicate that two genes which interact epistatically, one dominant and one recessive, are involved in the genetic control of leaf anthracnose resistance. As for stem anthracnose resistance, two genes also epistatic, one dominant and one recessive, explain the resistance to C. lindemuthianum race 83. The recessive gene is the same for leaf and stem resistance; however, the dominant genes are distinct and independent from each other. The three independent resistance genes of AN 910408 observed in this work could be derived from Guanajuato 31.

Lucerne biology and genetic improvement - an analysis of past activities and future goals in Australia

Breeding methodologies for cultivated lucerne (Medicago sativa L.), an autotetraploid, have changed little over the last 50 years, with reliance on polycross methods and recurrent phenotypic selection. There has been, however, an increase in our understanding of lucerne biology, in particular the genetic relationships between members of the M. sativa complex, as deduced by DNA analysis. Also, the differences in breeding behaviour and vigour of diploids versus autotetraploids, and the underlying genetic causes, are discussed in relation to lucerne improvement. Medicago falcata, a member of the M. sativa complex, has contributed substantially to lucerne improvement in North America, and its diverse genetics would appear to have been under-utilised in Australian programs over the last two decades, despite the reduced need for tolerance to freezing injury in Australian environments. Breeding of lucerne in Australia only commenced on a large scale in 1977, driven by an urgent need to introgress aphid resistance into adapted backgrounds. The release in the early 1980s of lucernes with multiple pest and disease resistance (aphids, Phytophthora, Colletotrichum) had a significant effect on increasing lucerne productivity and persistence in eastern Australia, with yield increases under high disease pressure of up to 300% being recorded over the predominant Australian cultivar, up to 1977, Hunter River. Since that period, irrigated lucerne yields have plateaued, highlighting the need to identify breeding objectives, technologies, and the germplasm that will create new opportunities for increasing performance. This review discusses major goals for lucerne improvement programs in Australia, and provides indications of the germplasm sources and technologies that are likely to deliver the desired outcomes.

Effects of phosphonate and salicylic acid treatments on anthracnose disease development and ripening of 'Kensington Pride' mango fruit

This study investigated treatment of mango (Mangifera indica L.) fruit with 2 host defence-promoting compounds for suppression of anthracnose disease (Colletotrichum gloeosporioides). Cultivar 'Kensington Pride' fruit were treated at concentrations of up to 1000 mg/L with either potassium phosphonate or salicylic acid. Applications were by various combinations of pre- and postharvest dips and vacuum infiltration. Postharvest treatments at up to 2000 mg/L salicylic acid were evaluated in a second fruiting season. Fruit were either uninoculated or inoculated with the fungal pathogen. Colour, firmness and disease-severity were assessed during shelf life at 23 degreesC. There were no significant (P>0.05) effects of potassium phosphonate or salicylic acid on anthracnose disease severity in the first season. Moreover, phosphonate or salicylic acid treatment did not significantly affect fruit colour or firmness changes. There were significant (P

Rootstocks influence 'Hass' avocado fruit quality and fruit minerals

To determine the potential for rootstocks to improve avocado quality, 'Hass' fruit from trees on seedling 'Velvick' (SV), clonal 'Velvick' (CV) or clonal 'Duke 7' (CD) rootstocks were harvested in 1999 and 2000, ripened at 20 C immediately after harvest (non-stored) or stored at 5 C for four weeks (stored), and then fruit quality and mineral concentrations determined. In 2000, non-stored CV fruit had lower severity of body rots (5% of flesh volume affected, caused mainly by Colletotrichum gloeosporioides) than CD fruit (20%), while body rots severity in stored CV fruit was 20% compared with 38% in stored CD fruit. Stored CV fruit had less severe diffuse discoloration (3% and 9% in 1999 and 2000, respectively) than CD fruit (7% and 20%, respectively). Stored CV fruit also had less severe vascular browning (19%) than CD fruit (33%) in 2000. In both seasons, CV fruit had 15-19% higher flesh calcium concentrations, 17-22% higher flesh boron concentrations, and 14-15% lower flesh nitrogen concentrations than CD fruit. In 2000, CV fruit also had 27% more skin calcium and 14% less skin nitrogen than CD fruit. There were no effects of rootstock on yield, canopy volume, measured length of non-suberized roots, average fruit weight, fruit length: width ratio, skin thickness, dry matter, proportion of the fruit skin purple-black when ripe, or seed mass: fruit weight ratio. These results suggest that there is potential to improve 'Hass' avocado quality through rootstock selection, and that tree and fruit mineral concentrations have a role in this relationship.

Ovary colonization by Claviceps africana is related to ergot resistance in male-sterile sorghum lines

Ergot, caused by Claviceps africana, has emerged as a serious threat to sorghum hybrid seed production worldwide. In the absence of gene-for-gene-based qualitative resistance in commercial cultivars, varieties with high pollen production that can escape ergot infection are preferred. Recent demonstration of differences in ergot susceptibility among male-sterile lines has indicated the presence of partial resistance. Using chitin-specific fluorescin-isothiocyanate-conjugated wheat germ agglutin and callose-specific aniline blue, this study investigated the process of sorghum ovary colonization by C. africana. Conidia germinated within 24 h after inoculation (a.i.); the pathogen was established in the ovary by 79 h a.i., and at least half of the ovary was converted into sphacelial tissue by 120 h a.i. Changes in fungal cell wall chitin content and strategic callose deposition in the host tissue were associated with penetration and invasion of the ovary. The rate of ovary colonization differed in three male-sterile lines that also differed in ergot susceptibility. This work demonstrates a possible histological basis for partial resistance in male-sterile sorghum lines that could lay the foundation for variety improvement through further breeding and selection.

Use of biofilm in the postharvest conservation of 'Pedro Sato' guava

This present work aimed to assess the effect of different concentrations of cassava starch, associated or not with prochloraz fungicide, on the postharvest conservation of 'Pedro Sato' guavas. Physiologically mature fruits were immersed in a solution of prochloraz (49.5 g/100 liters of water) for 5 min. Fruit treated with distilled water and air dried were used as control. They were immersed in cassava starch suspension at concentrations of 0, 20, 30 and 40 g/L, plus 0.5 mL/L of mineral oil. The fruits were stored at 21.0 ± 1.0 ºC and relative humidity of 85 ± 5%, for 12 days, and were examined at every three days. The combination of prochloraz and cassava starch resulted in delayed loss of firmness and yellowness and inhibited the incidence of lesions caused by Colletotrichum gloeosporioides during the 12 days of storage. Fruits treated with 40 g/L of starch, whether containing prochloraz or not, had unpleasant taste and odor, which suggests the occurrence of fermentation. Control fruits, with and without prochloraz, and those treated with starch with no prochloraz, had nearly 100% lesion occurrence within the 12 days. Starch suspension of 30 g/L, containing prochloraz, was the most effective in maintaining fruit quality.

Adubação nitrogenada e potássica na severidade da antracnose em dois cultivares de milho

O equilíbrio nutricional pode contribuir para a resistência das plantas às doenças. Com o objetivo de avaliar o efeito da interação entre as adubações nitrogenada e potássica na severidade das lesões da antracnose foliar e na nutrição mineral da cultura do milho, foram instalados dois experimentos, em casa de vegetação com dois cultivares, o DAS 2B710 (moderadamente resistente à doença) e o BRS 1010 (susceptível), cinco doses de N (75, 150, 300, 600 e 1200 mg dm-3) e cinco doses de K 63, 125, 250, 500 e 1000 mg dm-3). O delineamento experimental para cada cultivar foi em blocos inteiramente ao acaso, em esquema fatorial 5 x 5, com 25 tratamentos e quatro repetições. Cada vaso com quatro plantas constituiu uma parcela experimental. As doses de N e K foram divididas em quatro parcelas, com intervalos de dez dias a partir da semeadura, para os dois cultivares. Aos 21 dias após a semeadura, as plantas foram inoculadas e levadas para câmara úmida, com fotoperíodo de 16 horas, no escuro durante três dias consecutivos. Avaliou-se a severidade das lesões diariamente, até 11 dias após a inoculação. Aos 43 dias após a semeadura, a parte aérea das plantas foi colhida, secada e moída para determinar os teores de N e de K. A quantidade de área foliar lesionada dependeu da interação entre os nutrientes. Em ambos os cultivares, os menores valores de severidade foram observados na menor dose de N, combinada com a maior dose de K. A severidade das lesões observada no cultivar moderadamente resistente foi 41% menor que a observada no cultivar susceptível. A adubação nitrogenada influenciou de forma negativa o teor de K da parte aérea.

Severidade de doenças foliares e produtividade de genótipos de milho em resposta à adubação nitrogenada

As doenças foliares, principalmente, helmintosporiose (Bipolaris spp.), antracnose (Colletotrichum graminicola) e ferrugem (Puccinia spp.), podem ocasionar redução do rendimento da cultura do milho. Este estudo teve como objetivo avaliar a ocorrência e severidade de helmintosporiose, antracnose e ferrugem, bem como a produtividade de genótipos de milho, em resposta a doses crescentes de nitrogênio. Avaliou-se a severidade das doenças, utilizando-se escala de notas, em nove genótipos de milho (AG9040, AG7088, 30F35, 3F624, 1F583, 1F632, 1F557, 1F640 e ID219), submetidos a quatro doses de nitrogênio (67; 112; 157; e 202 kg ha-1), em duas safras (2009/2010 e 2010/2011). Para determinar a produtividade, utilizou-se a massa dos grãos coletados. Os graus de severidade da helmintosporiose, da antracnose e da ferrugem variaram em resposta às doses de nitrogênio e, entre os genótipos de milho avaliados durante as duas safras, entretanto, não houve relação clara entre as doses de nitrogênio aplicadas e a ocorrência das doenças nas duas safras. A ferrugem foi detectada apenas durante a safra I. Apesar da severidade das doenças foliares, elas, de modo geral, não afetaram significativamente a produtividade dos genótipos de milho avaliados. Desses, o genótipo de milho AG7088 pode ser considerado o mais produtivo e o mais resistente às doenças avaliadas.

Aplicação de extrato etanólico de própolis no crescimento e produtividade do feijoeiro comum

A própolis é uma substância muito utilizada em várias áreas da ciência, porém, os efeitos da sua aplicação em culturas anuais ainda são desconhecidos. Diante disto, objetivou-se, com este trabalho, verificar o efeito da aplicação de extrato etanólico de própolis (EEP) sobre a severidade da antracnose, o crescimento e a produtividade do feijoeiro carioca, cultivar IPR 139. Foram conduzidos dois experimentos no período de outubro de 2010 a janeiro de 2011. O delineamento experimental dos experimentos foi o de blocos ao acaso, com cinco repetições, realizando-se três aplicações de EEP nas concentrações: 0; 1; 2; 3 e 4%, quinzenalmente, a partir de 25 dias após a emergência (DAE). O EEP foi preparado com 10% de própolis bruta (originária do litoral paranaense) e 90% de álcool, a 96 ºGL. A altura das plantas, o número de folhas e a área foliar não foram alterados. O EEP na concentração de 4% reduziu a severidade da antracnose em até 63%, aumentou os teores foliares de nitrogênio (N), magnésio (Mg), ferro (Fe), a área foliar/folha e produtividade em até 33%. A redução da severidade da antracnose, obtida com a aplicação do EEP, aumentou a produtividade do feijoeiro.

Selection of anthracnose resistant common beans using detached leaves in partially controlled environment

The objectives of this study were to evaluate the possibility of selecting anthracnose resistant common bean plants using detached primary leaves in partially controlled environment of a greenhouse and identify differences in the reaction of genotypes to anthracnose. The common bean cultivars Ouro Negro, OuroVermelho, ManteigãoFosco 11, Rudá, Rudá-R, VP8, BRSMG Madrepérola, Pérola, MeiaNoite and BRSMG Talismãwere characterizedfor resistance to the races 65, 81 and 453 of Colletotrichum lindemuthianum and the method of detached primary leaves was compared to the method with the traditional inoculation of plants at the phenological stage V2. The lines Rudá, Rudá-R and Pérola were inoculated with the races 65 and 453 of C. lindemuthianum, aiming to assess the rate of coincidence of anthracnose severity by both inoculation methods. In general, the two methods presented similar results for the reaction of the cultivars. The use of detached primary leaves of common bean plants in the partially controlled environment was feasible for selection of plants resistant to anthracnose and has the advantages of low-needed infrastructure and reduction of resources, space and time.

Diversidade e prospecção de fungos endofíticos de Begonia spp. encontradas na Mata Atlântica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização da resistência à antracnose dos frutos verdes do cafeeiro

Dissertação apresentada para obtenção do Grau de Doutor em Biologia, especialidade Biologia Vegetal,pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Atividade antimicrobiana de fungos endofíticos isolados de plantas tóxicas da amazônia: Palicourea longiflora (aubl.) rich e Strychnos cogens bentham

Das plantas tóxicas da Amazônia Palicourea longiflora e Strychnos cogens foram isolados 571 fungos endofíticos e 74 bactérias endofíticas. Palicourea longiflora (Rubiaceae) e outras espécies desse gênero estão relacionadas a 90% das mortes de gado na região Amazônica. Strychnos cogens (Loganiaceae) e outras espécies de Strychnos são utilizadas pelos indígenas na confecção de "curares". Entre os endófitos isolados de P. longiflora foram identificados os fungos: Colletotrichum sp. e seu telemorfo Glomerella sp., Guignardia sp., Aspergillus niger, Phomopsis sp. e Xylaria sp., além da bactéria Burkholderia gladioli, pertencente a um grupo de fixadoras de nitrogênio. Dos isolados de S. cogens foram identificados os fungos: Colletotrichum sp., Guignardia sp., Aspergillus niger e Trichoderma sp. Uma amostra de 79 isolados fúngicos teve seus metabólitos extracelulares bioensaiados contra microrganismos patogênicos e fitopatogênicos: 19 isolados inibiram um ou mais microrganismos-teste. Dos oito isolados com melhores resultados de inibição, as móleculas bioativas eram menores que 12.000 daltons, fato verificado pela diálise dos metabólitos.

Tratamento térmico da manga: I - variação na temperatura e tempo de imersão

Estudou-se o efeito de tratamentos térmicos no controle das podridões da manga. Os tratamentos constaram na imersão de frutos de 'Haden' em água aquecida nas temperaturas de 45, 50, 55 e 60°C, usando-se para cada temperatura, 3 tempos de exposição 10, 20 e 30 minutos. Apos os tratamentos os frutos foram tratados com o ácido cloroetilfosfônico (Ethefon) a 500 ppm e o experimento foi encerrado no 10º dia após a execução dos tratamentos, com os frutos já maduros. Os melhores tratamentos foram aqueles a 50°C por 30 minutos e a 55°C por 10 minutos, resultando em controle praticamente total da antracnose, Colletotrichum gloeosporioides Penz, e não alteraram nem a aparência, nem os teores de ácidos e sólidos-solúveis dos frutos. Imersões em água a 55°C por 20 minutos e a 60°C por 10 minutos resultaram em escaldadura dos frutos.