What's in my Pot? Six Colletotrichum Species Causing Anthracnose in Brazilian Pecan Orchards.

Pecan (Carya illinoinensis) is one important exotic forest crop cultivated in South America, specifically in Brazil, Uruguay, and Argentina. However, diseases such as anthracnose, favored by high humidity conditions and high summer temperatures, make its cultivation difficult, causing important loss to pecan farmers. This study used morphological and molecular approaches to identify the Colletotrichum species causing anthracnose in pecan plantations in Southern Brazil. The isolates obtained from pecan fruits with anthracnose symptoms were grouped through quantitative morphological characteristics into three distinct morphotypes. Molecular analysis of nuclear genes allowed the identification of six species of Colletotrichum causing anthracnose in pecan: C. nymphaeae, C. fioriniae, C. gloeosporioides, C. siamense, C. kahawae, and C. karsti. Three of these species are reported for the first time as causal agents of anthracnose in pecan. Therefore, these results provide an important basis for the adoption and/or development of anthracnose management strategies in pecan orchards cultivated in southern Brazil and neighboring countries.

Old but still good: genetic diversity of ancient pecan genotypes from southern Brazil.

Pecan [Carya illinoinensis (Wangenh.) K. Koch] is a crop fruit native to the USA and Mexico currently cultivated in several countries, including Brazil, Uruguay, Argentina, Chile, Peru, China, South Africa, and Australia. Supported by the increasing consumption and market prices, the interest in the cultivation of this fruit crop is strongly growing around the world. In this study, AFLP and S-SAP markers were employed to characterize the genetic diversity of ancient accessions of pecan from southern Brazil. The evaluated plants were selected and preserved by the farmers and are remnants of the first introduction of seedlings from the U.S.A into southern Brazil aiming at developing research towards establishing commercial orchards. High levels of genetic diversity were estimated, suggesting that these plants have an important genetic background for the establishment of a germplasm collection with a wide genetic basis, for the development of breeding programs for this fruit crop. Cluster analysis of the genetic datasets revealed some correlation between the nuts' morphometric traits and genetic markers. Such correlation should be further exploited. These ancient genotypes must be evaluated for other agronomic traits of interest and included in core collections of pecans.

First report of Cercospora cf. malloti causing brown eyespot on strawberry in Brazil.

Strawberry (Fragaria x ananassa Duch.) is one of the most consumed fruits from the Rosaceae family in Brazil. It is cultivated on ~4,500 ha in all regions of the country, with a wide range of fungi causing leaf diseases. Strawberry leaves (cv. San Andreas) with symptoms of eyespot usually related to Mycosphaerella fragariae (Ramularia tulasnei) (Mazaro et al. 2006) were observed in April 2019, in a field located in Santa Maria, Rio Grande do Sul state, Brazil (29°43'19.6"S / 53°43'04.6"W). However, the symptoms slightly differed from Mycospherella blight as well as the fungi structures observed under an optical microscope. The symptomatic leaves had small circular spots, initially reddish-purple to dark brown color, which later increased in size. The center of the lesion was grayish pale, where dark brown conidiophores were found over it. The symptom occurred in 50% of the leaves of all plants during the crop season. To identify the causal agent, symptomatic leaves were collected and superficially disinfected with 70% ethanol for 30 s followed by 0.5% bleach for 30 s and then washed in sterile water. The leaves were placed in germination boxes with a moistened paper filter and incubated at 25 ºC and 12 h photoperiod for four days. After the formation of the fungal structures, the fungi was isolated in potato dextrose agar (PDA) medium and kept for 20 days at 25 ºC. Colonies were dark brown with pale pink aerial mycelium. The conidia were hyaline, single- to multi-septated with a filiform shape, straight or slightly curved. The fasciculate conidiophores on the stroma measured 20 to 70 x 2.5 to 3.5 µm. Conidia average measurement was 10.2 to 82.4 x 4. to 8.8 µm, similar to morphological characteristics of the genus Cercospora. The isolate named M6 was deposited in the Brazilian National System of Genetic Registration (SISGEN) no A57AE19. The DNA was extracted from a monosporic culture of M6 in PDA plates, which presented all typical aspects previously described. To confirm the identity, the PCR amplification was carried out for internal transcribed spacer (ITS) region ITS 1-5.8S - ITS 2 and Calmodulin region (CAL) genes (Acqua et al, 2011). The ITS (No. MZ713249) and CAL (No. MZ713254) sequences were deposited in the Genbank database. BLASTn analysis revealed the isolate was 99% similar to Cercospora cf. malloti which access codes in GenBank are KT 193689 and KT 193738 (Nguanhom et al. 2015). A pathogenicity test was performed to fulfill Koch's postulates. Strawberry plants with healthy leaves (60 days old), were inoculated by spraying a conidial suspension (1 × 105 conidia/ml). Control plants were sprayed with sterile water. The plants were incubated in a greenhouse at 28 ºC for 12 h photoperiod at 80% relative humidity under drip irrigation keeping at least 60% of field capacity. Six plants were used as replicates. Twenty days after inoculation, small circular reddish-purple spots were found, which later increased in size. Ten days later, the spots presented dark brown conidiophores over the center of the lesion. The symptoms described above were observed in all inoculated leaves. The control plants remained symptom-free. The pathogen was then re-isolated from the lesions and identified as C. cf. malloti based on the morphological aspects described above on PDA. In Thailand C. cf. malloti is reported on crops like Brassica alboglabra (cabbage) (Nguanhom et al. 2015), but not in strawberries. To our knowledge, this is the first report of C. cf. malloti pathogenic to strawberries in Brazil.

Diagrammatic scale for quantifying severity of brown leaf spot on Carya illinoinensis.

Brown leaf spot disease, caused by the fungus Ragnhildiana diffusa, is responsible for losses in pecan orchards and quantification is essential in order to conduct epidemiological studies to establish levels of productivity losses, and assess control strategies. Therefore, the aim of this study was to develop and validate a diagrammatic scale for assessing the severity of brown leaf spot in pecan. 240 symptomatic leaflets were collected from adult trees in Anta Gorda, in the Brazilian state of Rio Grande do Sul, and used to assess the current percentage severity of the disease. The scale was based on the minimum and maximum observed severity of the disease, with six graduations defined according to the Weber-Fechner Visual Stimulus Law. The scale therefore consisted of eight levels (1.5, 4.6, 10.6, 17.3, 39.5, 65.1, 77.5, and 98%). The estimates of eight evaluators provided a basis for validating the scale. They made estimates of disease severity on 40 symptomatic digitalized pecan leaflets first, without using the scale and then referring to it. The scale provided greater precision, accuracy, and reproducibility of estimates and should result in more reliable epidemiological studies.

Dormancy overcome and seedling quality of pecan in nursery / Superação de dormência e qualidade de mudas de nogueira-pecã em viveiro

ABSTRACT: This study evaluated the efficiency of methods to overcome seed dormancy in different storage periods in the production of pecan seedlings. Seeds were submitted to the following treatments: T1, T4 and T7 - control treatments (seeds with no treatment, stored at room temperature for 30, 60 and 90 days, respectively), T2, T5 and T8 - stratification (seeds were distributed in boxes with wet sand maintained at a temperature of 4°C for 30, 60 and 90 days, respectively), T3, T6 and T9 - scarification + stratification (seeds scarified with sandpaper n.80 and stratified by 30, 60 and 90 days, respectively), in completely random experimental design. Plant height, stem diameter, number of leaves, full emergence and emergence speed index (ESI) were evaluated after 14 weeks of sowing. The best development of pecan 'plants, their emergence, and ESI were observed in the stratification treatment for 90 day as well as in the scarification + stratification treatment for 90 day. Storing seeds in uncontrolled environment reduced their viability.

RESUMO: Neste trabalho, objetivou-se avaliar a eficiência dos métodos de superação de dormência de sementes, em diferentes períodos de armazenamento, na produção de mudas de nogueira-pecã. As sementes foram submetidas aos seguintes tratamentos: T1, T4 e T7 - testemunhas (sementes sem qualquer método de tratamento, armazenadas em ambiente não controlado por 30, 60 e 90 dias, respectivamente), T2, T5 e T8 - estratificação (sementes distribuídas em caixas com areia úmida e mantidas em temperatura de 4°C por 30, 60 e 90 dias, respectivamente), T3, T6 e T9 - escarificação + estratificação (sementes escarificadas com lixa n.80 e estratificadas também por 30, 60 e 90 dias, respectivamente), em delineamento experimental inteiramente ao acaso. Avaliou-se, após 14 semanas da semeadura, altura das plantas, diâmetro do colo, número de folhas, emergência total e índice de velocidade de emergência (IVE). O melhor desenvolvimento das plantas de nogueira-pecã, bem como sua emergência e IVE foi observado no tratamento com estratificação por 90 dias e quando combinado escarificação e estratificação por 90 dias. O armazenamento das sementes em ambiente não controlado reduz sua viabilidade.

Dimensionamento amostral para avaliação da massa e diâmetro de frutos de nogueira-pecã / Sample size for evaluating the weight and diameter of pecan fruits

O objetivo deste trabalho foi determinar o tamanho de amostra necessário para a estimação da média das massas do fruto, da casca e da amêndoa e dos diâmetros longitudinal e transversal de frutos de nogueira-pecã (Carya illinoinensis). Em 20 frutos, de cada uma de seis cultivares de nogueira-pecã ('Barton', 'Imperial', 'Importada', 'Mahan', 'Moneymaker' e 'Melhorada'), foi determinada a massa do fruto (MF), a massa da casca (MC) e a massa da amêndoa (MA) e mensurado o diâmetro longitudinal (DL) e o diâmetro transversal (DT). Com os dados desses 20 frutos, para cada caractere e cultivar, foram calculadas medidas de tendência central e de variabilidade, verificada a normalidade e calculado o tamanho de amostra. Para cada caractere, foi verificada a homogeneidade de variâncias entre as cultivares e comparadas as médias de cultivares. Cinquenta frutos são suficientes para a estimação da média de MF, MC, MA, DL e DT de frutos de nogueira-pecã, para erro de estimação de até 5% da média estimada, com grau de confiança de 95%.

The objective of this research was to determine the sample size necessary to estimate the average mass of fruit, peel and almond and longitudinal and transverse diameters of fruits of pecan (Carya illinoinensis). On 20 fruits of each of the six cultivars of pecan ('Barton', 'Imperial', 'Importada', 'Mahan', 'Moneymaker' e 'Melhorada'), the fruit mass (MF), the shell mass (MC) and almond mass (MA) was determined and measured the longitudinal diameter (DL) and transverse diameter (DT). With the data from these 20 fruits for each character and cultivar, measures of central tendency and variability were calculated, normality was checked and sample size was calculated. For each character, the homogeneity of variances among cultivars was verified and averages of cultivars were compared. Fifty fruits were enough to predict the average of MF, MC, MA, DL and DT fruit of pecan, with an estimation error equal to 5% of estimated average, with a degree confidence of 95%.