Assessment of renal and hepato-protective potential of guava leaves in male Sprague dawley rats.

Present research project was an attempt to explore the functional/nutraceutical worth of guava leaves from two locally grown varieties (Ruby & Safeda). For the purpose, guava leaves extract was fed to experimental male Sprague Dawley rats to explore the nutraceutical potential of guava leaves against hepatotoxicity. Two studies were performed on two types of rats i.e. study I (normal rats), study II (hepatotoxic rats). In both studies, 250 mg/kg each of pink guava leaves extracts (T1) and white guava leaves extracts (T2) was added in the feed. Feed intake and body weights of the rats were recorded. At the end of the first and eighth week of study, the blood samples of the rats were analyzed to check the effect of guava leaves extracts on renal functioning (Alkaline Phosphatase, Alanine Transaminase and Aspartate Transaminase) as well as liver functioning parameters including urea and creatinine. In both studies, comparatively higher feed consumption was observed in the control group than the rest of the treatments. At the end of study I, the highest weight (207±9.21 g) was observed in T0 whereas, during study II, the maximum value (202±5.58 g) was found in T2 (rats consuming white guava leaves extract) that indicates its effectiveness against hepatotoxicity. Regarding renal functioning tests, pink guava leaves were more effective in decreasing urea and creatinine levels in rats as compared to the white guava leaves in both study plans. Likewise, in each of study trial, pink guava leaves were more effective in reducing AST, ALT and ALP than white guava leaves and control. From the present investigation, it is deduced that guava leaves were effective against hepatotoxicity.

Development of novel g-SSR markers in guava (Psidium guajava L.) cv. Allahabad Safeda and their application in genetic diversity, population structure and cross species transferability studies.

Dearth of genomic resources particularly, microsatellite markers in nutritionally and commercially important fruit crop, guava necessitate the development of the novel genomic SSR markers through the library enrichment techniques. Three types of 3' -biotinylated oligonucleotide probes [(CT)14, (GT)12, and (AAC)8] were used to develop microsatellite enriched libraries. A total of 153 transformed colonies were screened of which 111 positive colonies were subjected for Sanger sequencing. The clones having more than five motif repeats were selected for primer designing and a total of 38 novel genomic simple sequence repeats could be identified. The g-SSRs had the motif groups ranging from monomer to pentamer out of which dimer group occurred the most (89.47%). Out of 38 g-SSRs markers developed, 26 were found polymorphic, which showed substantial genetic diversity among the guava genotypes including wild species. The average number of alleles per locus, major allele frequency, gene diversity, expected heterozygosity and polymorphic information content of 26 SSRs were 3.46, 0.56, 0.53, 0.29 and 0.46, respectively. The rate of cross-species transferability of the developed g-SSR loci varied from 38.46 to 80.77% among the studied wild Psidium species. Generation of N-J tree based on 26 SSRs grouped the 40 guava genotypes into six clades with two out-groups, the wild guava species showed genetic distinctness from cultivated genotypes. Furthermore, population structure analysis grouped the guava genotypes into three genetic groups, which were partly supported by PCoA and N-J tree. Further, AMOVA and PCoA deciphered high genetic diversity among the present set of guava genotypes including wild species. Thus, the developed novel g-SSRs were found efficient and informative for diversity and population structure analyses of the guava genotypes. These developed novel g-SSR loci would add to the new genomic resource in guava, which may be utilized in genomic-assisted guava breeding.

Genetic divergence among Psidium accessions based on single nucleotide polymorphisms developed for Eucalyptus.

The goal of this study was to analyze the genetic divergence among Psidium species accessions based on SNPs developed for Eucalyptus. Fifty-three Psidium accessions, including 47 P. guajava, were genotyped with EUCHIP60K. The dendrogram similarity ranged from 0.58 to 1.00, with a cophenetic value of 0.97. Five groups were identified at dendrogram cut point of 0.7: the first with 44 guava accessions, the second with 1 guava accession, the third with 3 P. guineense accessions, the forth with 2 guava accessions, and the fifth with 3 P. cattleianum accessions. The Bayesian analyses suggested seven subpopulations, with formation of two additional groups with guava accessions. Primers designed with Eucalyptus SNP sequences resulted in reliable Psidium amplicons on 6% polyacrylamide gels. In general, the SNP dendrogram agreed with biological genus structure, since different species were not grouped, indicating that transferability among Myrtaceae genus was possible and reliable.

Relationship between Psidium species (Myrtaceae) by resistance gene analog markers: focus on nematode resistance.

Guava (Psidium guajava L.) crop is severely affected by the nematode Meloidogyne enterolobii. Native Psidium species have been reported as sources of resistance against this nematode. Knowledge on the molecular relationship between Psidium species based on plant resistance gene analogs (RGA) can be useful in the genetic breeding of guava for resistance to M. enterolobii. In this study, RGA markers from conserved domains, and structural features of plant R genes, were employed to characterize Psidium species and establish genetic proximity, with a focus on nematode resistance. SSR markers were also applied owing to their neutral nature, thus differing from RGA markers. For this, species reported as sources of resistance to M. enterolobii, such as P. cattleianum and P. friedrichsthalianum, as well as species occurring in the Atlantic Rainforest and susceptible genotypes, were investigated. In 10 evaluated Psidium species, high interspecific genetic variability was verified through RGA and SSR markers, with intraspecific variation in P. guajava higher with SSR, as was expected. Resistant species were clustered by RGA markers, and differential amplicons among genotypes resistant and susceptible to M. enterolobii were identified. Knowledge on the molecular relationships between Psidium species constitutes useful information for breeding of the guava tree, providing direction for hybridization and material for rootstocks. Additionally, the genetic relationship between native species, which have been little studied, and P. guajava were estimated by RGAs, which were confirmed as important markers for genetic diversity related to pathogen resistance.

Estimativa da viabilidade polínica de araçazeiro (Psidium cattleianum Sabine) através de distintos métodos de coloração / Estimation of pollen viability of strawberry guava (Psidium cattleianum Sabine) through distinct staining methods

RESUMO A análise histoquímica dos grãos de pólen através do uso de corantes permite a determinação de sua viabilidade polínica. O objetivo deste estudo foi comparar a eficiência dos corantes orceína acética 2% e reativo de Alexander modificado, para estimar a viabilidade polínica de vinte acessos de araçazeiro (Psidium cattleianum Sabine) – uma árvore de frutos muito apreciados que também é utilizada para fins medicinais. Botões florais na pré-antese foram coletados e fixados em etanol:ácido acético (3:1) por 24 horas, após os botões foram transferidos para etanol 70% e mantidos sob refrigeração. A técnica de esmagamento das anteras foi utilizada no preparo das lâminas. Para cada acesso foram preparadas 2 lâminas por corante e analisados 500 grãos de pólen por lâmina. A viabilidade polínica foi estimada através da porcentagem de grãos de pólen viáveis. De modo geral, observou-se que os grãos de pólen corados com orceína acética 2% apresentaram viabilidade alta, acima de 98,1%, e os acessos não diferiram entre si. A viabilidade polínica através da coloração com a solução de Alexander variou de 43% (acesso Silveira Martins 1) a 97% (acesso Candelária 1), mostrando que provavelmente exista variabilidade genética entre os acessos. Em apenas dois acessos não houve diferença significativa entre os valores de viabilidade encontrados através dos dois corantes testados. Conclui-se então que a orceína acética 2% superestimou a viabilidade polínica de P. cattleianum e que o reativo de Alexander é o mais preciso, devido a sua dupla coloração (verde de malaquita + fucsina ácida). Assim, indica-se o reativo de Alexander para análise de estimativa da viabilidade polínica de genótipos de araçá envolvidos em um programa de melhoramento genético.

ABSTRACT The histochemical analysis of pollen grains through the use of stains makes it possible to determine their pollen viability. The purpose of this study was to compare the efficiency of 2% acetic orcein and Alexander’s stain modified, in estimating the pollen viability of twenty strawberry guava accessions (Psidium cattleianum Sabine) – a valued fruit tree which is used for medicinal purposes. Pre-anthesis flower buds were collected and fixed in ethanol:acetic acid (3:1 v/v) for 24 hours at room temperature, subsequently they were stored in 70% ethanol under refrigeration. The anther squash technique was performed for slide preparation. In each accession two slides per stain and 500 pollen grains were analyzed. The pollen viability was estimated according to the percentage of viable pollen grains. Overall, it was observed that pollen grains stained with 2% acetic orcein evinced a high viability, higher than 98,1%, and the accessions did not differ. The pollen viability estimated with Alexander’s stain varied from 43% (accession of Silveira Martins 1) to 97% (accession of Candelária 1), indicating that there is probably genetic variability among the accessions. There was no significant difference among the levels of viability found by means of the two stains in only two accessions. In conclusion, 2% acetic orcein overestimated the pollen viability of P. cattleianum, whereas the Alexander’s stain is in turn more accurate due to its two stains (green malachite and fuchsine). Thus, it is indicated Alexander’s stain for analysis estimation of pollen viability of strawberry guava genotypes involved in a genetic improvement program.

Avaliação da atividade antimicrobiana de extratos de Eugenia anomala e Psidium salutare (Myrtaceae) frente à Escherichia coli e Listeria monocytogenes / Evaluation of the antimicrobial activity of extracts of Eugenia anomala and Psidium salutare (Myrtaceae) against the Escherichia coli and Listeria monocytogenes

RESUMO As doenças transmitidas por alimentos ocorrem principalmente devido à ingestão de alimentos contaminados por microrganismos patogênicos, dentre eles a Escherichia coli e Listeria monocytogenes. Uma das alternativas estudadas para minimizar a contaminação de alimentos é o emprego de plantas, ou seus extratos, como agentes antimicrobianos de origem natural em produtos alimentícios. Desta forma o objetivo do presente estudo é fornecer dados científicos a respeito de duas plantas nativas do RS ainda não estudadas, Eugenia anomala e Psidium salutare, visando potencial emprego como agente antimicrobiano natural em alimentos. Para tanto, avaliou-se a atividade antimicrobiana de extratos de E. anomala e P. salutare contra E. coli e L. monocytogenes através da determinação da concentração inibitória mínima (CIM) pelo método de microdiluição em caldo, a capacidade antioxidante dos extratos por meio do método de redução do radical DPPH e a citotoxicidade in vitro empregando células CHO-K1. Os resultados obtidos mostraram que os extratos de acetato de etila e etanólico de ambas as espécies possuem ação antioxidante muito alta, de 94,08% e 93,86%, respectivamente. Apenas o extrato hexânico de P. salutare apresentou ação antimicrobiana moderada (CIM = 312,5 µg/mL). Todos os extratos apresentaram ação citotóxica sendo que os maiores percentuais foram do extrato clorofórmico de E. anomala (77,05%) e hexânico de P. salutare (76,79%), na concentração de 100 µg/mL. Assim, o presente estudo demonstrou que as espécies vegetais estudadas apresentam potencial para emprego como agente antimicrobiano destes microrganismos.

ABSTRACT The foodborne diseases occur mainly due to the ingestion of food contaminated by pathogenic microorganisms, including Escherichia coli and Listeria monocytogenes. One of the alternatives studied to minimize contamination of food is the use of plants or their extracts as antimicrobial agents naturally occurring in food products. The objective of this study is to provide scientific data on two native plants of RS have not studied Eugenia anomala and Psidium salutare for a potential use as a natural antimicrobial agent in food. To this end, we evaluated the antimicrobial activity of extracts of E. anomala and P. salutare against E. coli and L. monocytogenes by determining the minimum inhibitory concentration (MIC) by the broth microdilution method, the antioxidant capacity of the extract for means DPPH radical reduction method and in vitro cytotoxicity using CHO-K1 cells. The results showed that the ethyl acetate and ethanolic extracts of both species have very high antioxidant activity, of 94.08% and 93.86%, respectively. Only the hexane extract of P. salutare showed a moderate antimicrobial activity (MIC = 312.5 mg/mL). Moreover, all extracts showed cytotoxic action of which the highest percentages were the chloroform extract of E. anomala (77.05%) and hexane P. salutare (76.79%) at a concentration of 100 mg/mL. Thus, the present study showed that plant species have potential for use as an antimicrobial agent against these microorganisms.

SSR markers: a tool for species identification in Psidium (Myrtaceae).

Molecular DNA markers are used for detection of polymorphisms in individuals. As they are independent of developmental stage of the plant and environmental influences, they can be useful tools in taxonomy. The alleles of simple sequence repeat (SSR) markers (or microsatellites) are traditionally used to identify taxonomic units. This application demands the laborious and costly delimitation of exclusive alleles in order to avoid homoplasy. Here, we propose a method for identification of species based on the amplification profile of groups of SSR markers obtained by a transferability study. The approach considers that the SSR are conserved among related species. In this context, using Psidium as a model, 141 SSR markers developed for Psidium guajava were transferred to 13 indigenous species of Psidium from the Atlantic Rainforest. Transferability of the markers was high and 28 SSR were conserved in all species. Four SSR groups were defined and they can help in the identification of all 13 Psidium species studied. A group of 31 SSR was genotyped, with one to six alleles each. The H0 varied from 0.0 to 0.46, and PIC from 0.0 to 0.74. Cluster analysis revealed shared alleles among species. The high percentage of SSR transferability found in Psidium evidences the narrow phylogenetic relationship existing among these species since transferability occurs by the preservation of the microsatellites and anchoring regions. The proposed method was useful for distinguishing the species of Psidium, being useful in taxonomic studies.

Protective effect of guabiju (Myrcianthes pungens (O. Berg) D. Legrand) and red guava (Psidium cattleyanum Sabine) against cisplatin-induced hypercholesterolemia in rats

The present study was the first to examine the protective effect and antioxidant potential of two fruits native to the south of Brazil, namely guabiju (Myrcianthes pungens (O. Berg) D. Legrand) and red guava (Psidium cattleyanum Sabine), with respect to reducing the effects of cisplatin-induced hypercholesterolemia in male Wistar rats. The cisplatin-treated animals showed high glucose, LDL cholesterol, oxidized LDL cholesterol and total cholesterol levels. These native fruits reversed the increase in these parameters. The fat levels in the liver improved with supplementation with the fruits, and there were no differences in the thiobarbituric acid reactive substances (TBARS), creatinine, urea or HDL cholesterol levels, nor in the weight of the organs analyzed, indicating that the dose used did not affect these markers.

O presente estudo foi o primeiro realizado para avaliar o efeito protetor e o potencial antioxidante de duas frutas nativas do sul do Brasil, conhecidas como Guabiju (Myrcianthes pungens (O. Berg) D. Legrand) e araçá vermelho (Psidium cattleyanum Sabine), em relação a reduzir os efeitos da hipercolesterolemia induzida pela cisplatina em ratos Wistar machos. Os animais tratados com a cisplatina apresentaram elevada taxa de glicose, altos níveis de colesterol LDL, colesterol LDL oxidado e colesterol total. Os frutos nativos atuaram protegendo os animais pela redução destes parâmetros. Os níveis de gordura no fígado melhoraram com a suplementação pelos frutos e não houve diferenças nas substâncias reativas ao ácido tiobarbitúrico (TBARS) e níveis de creatina, uréia e colesterol HDL e, também, no peso dos órgãos analisados, indicando que a dose utilizada não foi capaz de afetar tais marcadores.

Allelic database and divergence among Psidium accessions by using microsatellite markers.

This study aimed to investigate the genetic variability among guava accessions and wild Psidium species of the Embrapa Semiárido germplasm collection by using microsatellite loci to guide genetic resources and breeding programs, emphasizing crosses between guava and other Psidium species. DNA was extracted using the 2X CTAB method, and polymerase chain reaction products were analyzed on 6% denatured polyacrylamide gels stained with silver nitrate. The unweighted pair-group method using arithmetic average dendrogram generated from the distance matrix of the Jaccard coefficient for 183 alleles of 13 microsatellite loci was used for visualization of genetic similarity. The number of base pairs was estimated using inverse mobility method based on the regression of known-size products. Analysis of molecular variance was performed using total decomposition between and within guava accessions. The accessions showed similarity from 0.75 to 1.00, with the dendrogram presenting cophenetic value of 0.85. Five groups were observed: the first included guava accessions; the second, P. guineense accessions; the third, one accession of P. friedrichsthalianum; and the last 2 groups, P. cattleianum. The genetic similarity among P. guineense and some guava accessions were above 80%, suggesting greater possibility to obtain interspecies hybrids between these 2 species. The genetic variability between the accessions was considered to be high (ΦST = 0.238), indicating that guava genetic variability is not uniformly distributed among the 9 Brazilian states from where the accession were obtained. Obtaining a greater number of accessions by Brazilian states is recommended in order to have greater diversity among the species.

Nutritional and nutraceutical comparison of Jamaican Psidium cattleianum (strawberry guava) and Psidium guajava (common guava) fruits.

Psidium cattleianum (strawberry guava) is one of many underutilised edible fruits that grow wild in Jamaica, and could potentially be commercially exploited to yield health and economic benefits. In this study, the total phenolics, proximate contents, and antioxidant, anti-inflammatory, and antimicrobial activities of P. cattleianum and P. guajava (common guava), a well-known species, were compared. Strawberry guavas were found to be superior to common guavas in antioxidant and antimicrobial activities, total phenolics and vitamin C content. They also possessed relatively high fibre content (24.9%). The hexane and ethyl acetate extracts of strawberry guavas showed cyclooxygenase-2 enzyme inhibitory activities of 18.3% and 26.5%, respectively (250 µg/mL), indicating anti-inflammatory activity. The EtOAc and MeOH extracts of P. guajava showed 56.4% (COX-2) and 44.1% (COX-1) inhibitory activity, respectively. Additionally, nine compounds were isolated from strawberry guava fruits, some of which demonstrated anti-inflammatory activity. These results indicate that strawberry guavas are beneficial for health.

Aroma volatile compositions of high- and low-aromatic guava varieties.

BACKGROUND: The traditional guava variety cultivated in Israel, 'Ben Dov', emits a very strong odour, whereas some newly bred varieties have a mild odour. In this study the aroma profile composition of the high-aromatic 'Ben Dov' variety was compared with those of four new low-aromatic varieties. RESULTS: Overall, using gas chromatography/mass spectrometry, a total of 30 aroma volatiles were detected in fresh ripe guava fruit: 15 of them were specifically detected only in the high-aromatic 'Ben Dov' variety, 13 were detected in both the high- and low-aromatic varieties and two were detected only in the new low-aromatic varieties. Interestingly, 11 out of the 15 volatiles specifically detected in 'Ben Dov' were esters that contribute sweet, tropical and fruity notes. In contrast, ten out of 13 detected terpenes and two detected aldehydes, contributing green, spicy, herbal and woody notes, were common to both the high- and low-aromatic varieties. CONCLUSION: Based on these findings, it is concluded that accumulation of esters is the main reason why the traditional 'Ben Dov' guava variety emits such a strong tropical fruity odour. In contrast, the newly bred low-aromatic guava varieties did not synthesise esters at all and thus lacked fruity aromatic notes. Overall, the results of this study point out the important role of esters in forming tropical fruity guava odours.

Dimorfismo sexual do gorgulho-da-goiaba Conotrachelus psidii Marshall (Coleoptera: Curculionidae) / Sexual dimorphism of guava weevil Conotrachelus psidii Marshall (Coleoptera: Curculionidae)

Este trabalho teve como objetivo determinar diferenças que permitam identificar o sexo de adultos de Conotrachelus psidii Marshall, 1922. Gorgulhos coletados em um pomar comercial de goiaba foram individualizados em caixas e mantidos em laboratório. Diariamente, dez gorgulhos eram agrupados e seu comportamento observado. Durante o acasalamento, os machos e as fêmeas eram identificados. Cinqüenta gorgulhos de cada sexo foram sacrificados e observados sob microscópio estereoscópico. Foram examinadas a inserção da antena no rostro e a distribuição de pilosidade na superfície do tórax e abdome. Também, foi medido o comprimento e a largura do corpo, pronoto, último esternito abdominal e rostro. Do ponto de vista comportamental, machos emitem sons por estridulação enquanto que as fêmeas não. A largura do corpo, do pronoto e o comprimento do corpo e do rostro nas fêmeas foram maiores, enquanto que a largura do último esternito abdominal foi menor. Três outras diferenças morfológicas são visíveis a olho nu: 1) A região anterior do pronoto das fêmeas possui pilosidade reduzida ou ausente enquanto que nos machos é abundante; 2) o último tergito nas fêmeas fica recoberto pelo penúltimo enquanto que nos machos é parcialmente visível; 3) o primeiro esternito abdominal nas fêmeas é mais proeminente, convexo e com pilosidade reduzida ou ausente. Nos machos esse segmento é ligeiramente côncavo e com pilosidade em toda a superfície. As diferenças entre machos e fêmeas encontradas neste trabalho permitem, sem danificar os insetos nem alterar seu comportamento, uma sexagem eficiente do gorgulho-da-goiaba.

We investigate structural and behavioral differences between male and female adults of guava weevil, Conotrachelus psidii Marshall, 1922 to help in sex determination. Weevils collected from a commercial guava orchard were individually caged and maintained under laboratory conditions. Every day ten individuals were grouped and their behavior was observed. During mating the males and females were identified. Fifty weevils of each sex were killed and they were observed with stereoscopic microscope. The antenna insertion on the rostrum and hair on the thorax and abdomen surfaces were examined. Length and width of body, pronotum, last abdominal sternite and rostrum were also taken. Behaviorally, the male weevils produce audible sound by stridulation of abdomen while females do not. In the females, the body and pronotum width and body a rostrum length were higher than in the males, but the last abdominal sternite was smaller. Three other structural differences were visible with the naked eye: 1) the anterior region of pronotum have scarce hair or have not in the females and hair is dense in the males; 2) the last tergite of females is hidden by the penultimate tergite whereas in the males is visible; 3) the first abdominal sternite of females is more prominent, very convex and without or with little bristle. In the males it is slightly concave and with abundant hair. The male-female differences found in this work allow, without hurt or change insect behavior, an efficient sex determination of guava weevil.