Carica papaya L. sex chromosome review and physical mapping of the serk 2, svp-like and mdar 4 sequences.

Physical mapping evidences the chromosome organization and structure. Despite the data about plant cytogenomics, physical mapping has been conducted from single-copy and/or low-copy genes for few species. Carica papaya cytogenomics has been accomplished from BAC-FISH and repeatome sequences. We aimed to map the serk 2, svp-like and mdar 4 sequences in C. papaya. The sequences were amplified and the amplicons sequenced, showing similarity in relation to serk 2, svp-like and mdar 4 genes. Carica papaya diploidy was confirmed and the mitotic chromosomes characterized. The chromosome 1 exhibited the secondary constriction pericentromeric to the centromere of the long arm. So, we concluded that it is the sex chromosomes. serk 2 was mapped in the long arm interstitial portion of the sex chromosomes, and the interphase nuclei showed two fluorescence signals. Considering these results and the sequencing data from the C. papaya sex chromosomes, svp-like and mdar 4 genes were mapped in the interstitial region of the sex chromosome long arm. Both sequences showed only one fluorescence signal in the interphase nuclei. The procedure adopted here can be reproduced for other single-copy and/or low-copy genes, allowing the construction of cytogenetic maps. In addition, we revisited the cytogenomics data about C. papaya sex chromosomes, presenting a revised point of view about the structure and evolution to these chromosomes.

Popcorn (Zea mays L. var. Everta) haploids identified by Navajo phenotype and ploidy level.

For popcorn, obtaining and identifying haploids are still challenging steps. We aimed to induce and screen haploids in popcorn using the Navajo phenotype, seedling vigor, and ploidy level. We used the Krasnodar Haploid Inducer (KHI) in crosses with 20 popcorn source germplasms and five maize controls. The field trial design was completely randomized, with three replications. We assessed the efficacy of induction and identification of haploids based on the haploidy induction rate (HIR) and false positive and negative rates (FPR and FNR). Additionally, we also measured the penetrance of the Navajo marker gene (R1-nj). All putative haploids classified by the R1-nj were germinated together with a diploid sample and evaluated for false positives and negatives based on vigor. Seedlings from 14 females were submitted to flow cytometry to determine the ploidy level. The HIR and penetrance were analyzed by fitting a generalized linear model with a logit link function. The HIR of the KHI, adjusted by cytometry, ranged from 0.0 to 1.2%, with a mean of 0.34%. The average FPR from screening based on the Navajo phenotype was 26.2% and 76.4% for vigor and ploidy, respectively. The FNR was zero. The penetrance of R1-nj ranged from 30.8 to 98.6%. The average number of seeds per ear in temperate germplasm (76) was lower than that obtained in tropical germplasm (98). There is an induction of haploids in germplasm of tropical and temperate origin. We recommend the selection of haploids associated with the Navajo phenotype with a direct method of confirming the ploidy level, such as flow cytometry. We also show that haploid screening based on Navajo phenotype and seedling vigor reduces misclassification. The origin and genetic background of the source germplasm influence the R1-nj penetrance. Because the known inducers are maize, developing doubled haploid technology for popcorn hybrid breeding requires overcoming unilateral cross-incompatibility.

In Vitro Polyploidization of Brassolaeliocattleya Hybrid Orchid.

The Cattleya (Orchidaceae-Laeliinae subtribe) intergeneric hybrids, such as Brassolaeliocattleya (Blc.), have great ornamental value, due to their compact-size, with large and high color diversity of flowers. Artificial induction of polyploidy brings agronomic, ornamental and genetic benefits to plants. Polyploidization efficiency depends on factors, such as the type of antimitotic, polyploidization method, concentrations, exposure times and type of explant. This study aimed to develop a protocol to polyploidize Blc. orchids, by testing two types of explants (seeds and protocorms), concentrations and exposure times to colchicine. The effects of colchicine on the in vitro development of explants were also investigated. The responses of explants to colchicine depended on the concentrations, exposure time and the interaction of these factors. Flow cytometric analysis evidenced high endopolyploidy and allowed the separation of polyploidized (4C, 8C and 16C peaks) from non-polyploidized (only 2C and 4C peaks) plants. The highest percentage of polyploid plants was regenerated from protocorms (16.4%) treated with colchicine instead of seeds (3.2%). Protocorms treated with colchicine at 500-750 µM for 18 h resulted in the best percentage of polyploidization. Additionally, in vitro natural polyploidization using protocorms was reported (11.5%). Cytological analyses allowed an estimation of the number of chromosomes of the parents (≡70), polyploidized (≡140) and non-polyploidized progeny (≡70).

Cadmium-mediated toxicity in plant cells is associated with the DCD/NRP-mediated cell death response.

Cadmium (Cd2+ ) is highly harmful to plant growth. Although Cd2+ induces programmed cell death (PCD) in plant cells, Cd2+ stress in whole plants during later developmental stages and the mechanism underlying Cd2+ -mediated toxicity are poorly understood. Here, we showed that Cd2+ limits plant growth, causes intense redness in leaf vein, leaf yellowing, and chlorosis during the R1 reproductive stage of soybean (Glycine max). These symptoms were associated with Cd2+ -induced PCD, as Cd2+ -stressed soybean leaves displayed decreased number of nuclei, enhanced cell death, DNA damage, and caspase 1 activity compared to unstressed leaves. Accordingly, Cd2+ -induced NRPs, GmNAC81, GmNAC30 and VPE, the DCD/NRP-mediated cell death signalling components, which execute PCD via caspase 1-like VPE activity. Furthermore, overexpression of the positive regulator of this cell death signalling GmNAC81 enhanced sensitivity to Cd2+ stress and intensified the hallmarks of Cd2+ -mediated PCD. GmNAC81 overexpression enhanced Cd2+ -induced H2 O2 production, cell death, DNA damage, and caspase-1-like VPE expression. Conversely, BiP overexpression negatively regulated the NRPs/GmNACs/VPE signalling module, conferred tolerance to Cd2+ stress and reduced Cd2+ -mediated cell death. Collectively, our data indicate that Cd2+ induces PCD in plants via activation of the NRP/GmNAC/VPE regulatory circuit that links developmentally and stress-induced cell death.

Repetitive sequences and structural chromosome alterations promote intraspecific variations in Zea mays L. karyotype.

LTR-retrotransposons, knobs and structural chromosome alterations contribute to shape the structure and organization of the Zea mays karyotype. Our initial nuclear DNA content data of Z. mays accessions revealed an intraspecific variation (2 C = 2.00 pg to 2 C = 6.10 pg), suggesting differences in their karyotypes. We aimed to compare the karyotypes of three Z. mays accessions in search of the differences and similarities among them. Karyotype divergences were demonstrated among the accessions, despite their common chromosome number (2n = 20) and ancestral origin. Cytogenomic analyses showed that repetitive sequences and structural chromosome alterations play a significant role in promoting intraspecific nuclear DNA content variation. In addition, heterozygous terminal deletion in chromosome 3 was pointed out as a cause of lower nuclear 2 C value. Besides this, translocation was also observed in the short arm of chromosome 1. Differently, higher 2 C value was associated with the more abundant distribution of LTR-retrotransposons from the family Grande in the karyotype. Moreover, heteromorphism involving the number and position of the 180-bp knob sequence was found among the accessions. Taken together, we provide insights on the pivotal role played by repetitive sequences and structural chromosome alterations in shaping the karyotype of Z. mays.

Author Correction: Repetitive sequences and structural chromosome alterations promote intraspecific variations in Zea mays L. karyotype.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Plant Chromosome-Specific Probes by Microdissection of a Single Chromosome: Is That a Reality?

Painting plant chromosomes through chromosomal in situ suppression (CISS) hybridization has long been considered impracticable. Seeking to build specific and complex probes from a single microdissected chromosome, we employed human chromosomes as models to standardize all the necessary steps for application in plants. Human metaphases were used to define the adequate conditions for microdissection, chromosome DNA amplification and labeling through degenerate oligonucleotide-primed PCR, and in situ hybridization stringency. Subsequently, these methodologies were applied in the plant species Zea mays (chromosome 1) and Capsicum annuum (chromosome 7 or 8). The high quality of human and plant cytogenetic preparations and the meticulous standardization of each step, especially the most critical ones - microdissection and first round of DNA amplification - were crucial to eliminate the signs of non-specific hybridization and for direct application in plants. By overcoming these challenges, we obtained chromosome-specific probes, which allowed to achieve a clear and uniform painting of the entire target chromosomes with little or no background, evidencing their complexity and specificity. Despite the high amount of ubiquitous repetitive sequences in plant genomes, the main drawback for chromosome painting, we successfully employed our methodology on two plant species. Both have more than 80% repetitive sequences, which is compared to the human genome (66-69%). This is the first time that plant chromosome-specific probes were successfully obtained from a single A mitotic or meiotic microdissected chromosome. Thereby, we assume that chromosome painting through microdissection and CISS hybridization can now be considered a reality in the field of plant cytogenetics.

Updating the maize karyotype by chromosome DNA sizing.

The karyotype is a basic concept regarding the genome, fundamentally described by the number and morphological features of all chromosomes. Chromosome class, centromeric index, intra- and interchromosomal asymmetry index, and constriction localization are important in clinical, systematic and evolutionary approaches. In spite of the advances in karyotype characterization made over the last years, new data about the chromosomes can be generated from quantitative methods, such as image cytometry. Therefore, using Zea mays L., this study aimed to update the species' karyotype by supplementing information on chromosome DNA sizing. After adjustment of the procedures, chromosome morphometry and class as well as knob localization enabled describing the Z. mays karyotype. In addition, applying image cytometry, DNA sizing was unprecedentedly measured for the arms and satellite of all chromosomes. This way, unambiguous identification of the chromosome pairs, and hence the assembly of 51 karyograms, were only possible after the DNA sizing of each chromosome, their arms and satellite portions. These accurate, quantitative and reproducible data also enabled determining the distribution and variation of DNA content in each chromosome. From this, a correlation between DNA amount and total chromosome length evidenced that the mean DNA content of chromosome 9 was higher than that of chromosome 8. The chromosomal DNA sizing updated the Z. mays karyotype, providing insights into its dynamic genome with regards to the organization of the ten chromosomes and their respective portions. Considering the results and the relevance of cytogenetics in the current scenario of comparative sequencing and genomics, chromosomal DNA sizing should be incorporated as an additional parameter for karyotype definition. Based on this study, it can be affirmed that cytogenetic approaches go beyond the simple morphological description of chromosomes.

Study of oviposition behavior of aedes aegypti in two neighborhoods under the influence of semi-arid climate in the municipality of salinas, state of Minas Gerais, Brazil

The mosquito Aedes aegypti (Diptera: Culicidae) is the main vector of human arborvirus in Brazil.Oviposition behavior is important for the survival and dispersal of A. aegypti. The objective of this studywas to evaluate the oviposition behavior and its relationship to meteorological factors. Twenty ovitrapswere installed in two neighborhoods in the urban area of the city of Salinas, MG, southeastern Brazil.The traps were inspected weekly for the presence of eggs in the water and on the paddle. Simultaneously,regional weather data were analyzed. The results showed the Ovitrap Positivity Index (OPI) variedgreatly, from 0 to 90 percent, between the weeks and boroughs sampled, demonstrating the vector’s greatpopulation fluctuation. Among the analyzed climatic factors, rainfall strongly influenced oviposition,followed by relative air humidity. The number of eggs found on the paddle was significantly higher thanin the water (p<0.05). However, a significant increase of eggs in the water was noticed when the relativehumidity dropped below 40 percent. This seems to be an important behavior to maintain the population inperiods of drought. Through the low number of eggs (<30) found in 56 percent of positive traps, we concludedthat skip-oviposition behavior occurred, contributing to the prevalence of mosquitoes during thesampled period. It is concluded that the occurrence of water oviposition may be higher than described inthe literature, and should be studied due to its evolutionary and epidemiological importance...

O mosquito Aedes aegypti é o principal vetor de arborvírus humanos no Brasil. O comportamentode oviposição é importante para a sobrevivência e dispersão do A. aegypti. O objetivo deste trabalho foi avaliar estes comportamentos e sua relação com fatores climáticos. Instalaram-se vintearmadilhas ovitrampa em dois bairros de Salinas-MG. Todas foram inspecionadas semanalmentepara contagem e identificação dos ovos presentes na água e na palheta. Paralelamente, analisaram-sedados meteorológicos da região. Os resultados mostraram que o Índice de Positividade deOvitrampa (IPO) variou muito (0 por cento a 90 por cento) entre as semanas/bairros amostrados, o que demonstraa grande flutuabilidade populacional do vetor. Dentre os fatores climáticos analisados, o que maisintensamente influenciou a oviposição foi a precipitação, seguida da umidade relativa. O númerode ovos encontrados na palheta foi significativamente maior que na água (p<0,05). Entretanto,observou-se aumento significativo de ovos na água quando a umidade relativa esteve abaixo de 40 por cento.Este parece ser um dado importante para a manutenção das populações em períodos de seca. O baixonúmero de ovos (<30) encontrados em 56 por cento das armadilhas positivas demonstrou a ocorrência docomportamento de skip-oviposition que pareceu contribuir para a prevalência do mosquito em todoo período amostrado. Concluiu-se que a ocorrência de oviposição na água pode ser maior do que odescrito na literatura e deve ser investigada em virtude de sua importância evolutiva e epidemiológica...