Specific Physiological and Anatomical Traits Associated With Polyploidy and Better Detoxification Processes Contribute to Improved Huanglongbing Tolerance of the Persian Lime Compared With the Mexican Lime.

Huanglongbing (HLB) is presently a major threat to the citrus industry. Because of this disease, millions of trees are currently dying worldwide. The putative causal agent is a motile bacteria belonging to Candidatus Liberibacter spp., which is transmitted by psyllids. The bacteria is responsible for the synthesis of callose at the phloem sieve plate, leading to the obstruction of the pores that provide connections between adjacent sieve elements, thus limiting the symplastic transport of the sugars and starches synthesized in leaves to the other plant organs. The Persian triploid lime (Citrus latifolia) is one of the most HLB-tolerant citrus varieties, but the determinants associated with the tolerance are still unknown. HLB-infected diploid Mexican lime (Citrus aurantiifolia) and Persian lime were investigated. The leaf petiole was analyzed using scanning electron microscopy (SEM) to observe callose deposition at the phloem sieve plate. Leaf starch contents and detoxification enzyme activities were investigated. In the field, Persian lime leaves present more limited symptoms due to HLB than the Mexican lime leaves do. Photosynthesis, stomatal conductance, and transpiration decreased compared with control plants, but values remained greater in the Persian than in the Mexican lime. Analysis of the petiole sieve plate in control petiole samples showed that pores were approximately 1.8-fold larger in the Persian than in the Mexican lime. SEM analyses of petiole samples of symptomatic leaves showed the important deposition of callose into pores of Mexican and Persian limes, whereas biochemical analyses revealed better detoxification in Persian limes than in Mexican limes. Moreover, SEM analyses of infected petiole samples of asymptomatic leaves showed much larger callose depositions into the Mexican lime pores than in the Persian lime pores, whereas biochemical traits revealed much better behavior in Persian limes than in Mexican limes. Our results reveal that polyploids present specific behaviors associated with important physiological and biochemical determinants that may explain the better tolerance of the Persian lime against HLB compared with the Mexican lime.

Return temperature after heat shock affects the production of tetraploids in the yellowtail tetra Astyanax altiparanae.

The aim of this study was to evaluate different post-shock temperatures for tetraploid induction in the yellowtail tetra Astyanax altiparanae. Newly fertilized eggs were divided into four groups, three were submitted to heat shock (40°C for 2 min) at 24 min post-fertilization (mpf) and another group remained without shock (control). Groups submitted to temperature shock were further separated at the following temperatures: 22°C, 26°C and 28°C. Survival among embryonic development was counted and at hatching the ploidy was analyzed by flow cytometry. The results showed that the post-shock temperature affects the parameters analyzed and, therefore, must be considered for optimization of the production of tetraploid in A. altiparanae. Those data are innovative and could be used in future studies of basic biology in this species.

Secondary origin, hybridization and sexual reproduction in a diploid-tetraploid contact zone of the facultatively apomictic orchid Zygopetalum mackayi.

The production of triploids and apomictic reproduction are important processes for polyploid establishment and cytotype coexistence, but we know little about the interaction between triploids and facultatively apomictic plants. To bridge this gap, we studied the pollen-dependent, facultatively apomictic orchid Zygopetalum mackayi from high-elevation outcrops of southeast Brazil. We described the nature of the contact between Z. mackayi cytotypes and patterns of genetic diversity and structure based on eight microsatellite markers and 155 individuals of pure tetraploid, pure diploid and mixed cytotype populations. Our results revealed high values of genetic and genotypic diversity within all populations of Z. mackayi. Each cytotype emerged as a genetic distinct cluster, combining individuals from different populations. Triploids clustered in an intermediate position between diploids and tetraploids. Most genetic variance is associated with individuals within populations and genetic differentiation is high among populations. Mixed cytotype populations of Z. mackayi originate from secondary contact. Triploids are hybrids between diploids and tetraploids and likely act as a bridge. Our results point to the predominance of sexual reproduction in all populations but do not corroborate previous basic chromosome number for this species. Polyploidy rather than facultative apomixis may explain the larger geographic distribution of tetraploids of Z. mackayi.

In vivo storage of oocytes leads to lower survival, increased abnormalities and may affect the ploidy status in the yellowtail tetra Astyanax altiparanae.

SummaryIn this study we analyzed whether the in vivo storage of oocytes (time after ovulation until fertilization) affects the survival and the ploidy status of the yellowtail tetra Astyanax altiparanae. Fish were induced to spawn and, after ovulation, a small aliquot was stripped and immediately fertilized (positive control group). Subsequently, aliquots (~150 oocytes) were stripped and fertilized at various time points of 60, 120, 180 or 240 min. Developmental stages, abnormalities, survival and the ploidy status of the hatched larvae were examined. As expected, in the control group, 100% of the larvae were diploid. Conversely, triploid individuals were observed just at the 60 min treatment time point (0.6%). In vivo storage of oocytes also influenced the survival rates (P < 0.05); the 180 and 240 min samples, respectively, presented lower survival rates at gastrula (50.10±6.26% and 40.92±5.32%), and somite (17.80±5.14% and 4.41±2.76%) stages and lower hatching rates (12.01±4.04% and 4.41±2.76%). A higher percentage (99.27±0.40%) of normal larvae and only a few abnormal larvae (0.73±0.40%) were observed in the control group (P = 0.0000). This observation did not differ from that observed at the 60 min treatment point (P = 0.9976). A significant increase in the percentage of abnormalities was observed in the other treatments, and, after 240 min, the highest percentage of abnormal larvae was seen (P=0.0024; 83.33±16.67%). In conclusion, we showed that oocyte ageing had a significant effect on survival and may affect the ploidy status in A. atiparanae.

Triploidy in the hematology of jundia juveniles (Siluriformes: Heptapteridae) / Triploidia na hematologia de juvenis de jundiá (Siluriformes: Heptapteridae)

This study compared the hematological characteristics of diploid and triploid of jundia, Rhamdia quelen juveniles, an important freshwater fish cultured in south Brazil. Hematological morphometry of erythrocytes were determined in blood smears under a light microscope. The blood was used to measure the number of red blood cells (RBC) with a hemocytometer Neubauer chamber, and the numbers of white blood cells (WBC) and thrombocytes that were obtained using an indirect method. The results showed that triploidy increased (p < 0.01) the size and volume of the erythrocytes. Nevertheless, as expected, triploidy decreased (p < 0.01) the number of circulating erythrocytes, leucocytes and trombocytes in the blood of jundia. Moreover differential cell counts were different in diploids and triploids, suggesting that triploidy affects the number of cells and their proportion in blood. Lymphocytes were the most predominant cells in the differential counting of diploid fish (62.5 percent) while monocytes were predominant in triploid fish (49.6 percent). These results suggest performance differences between ploidies of jundia, and require future studies to evaluate the potential of triploid jundia in the culture conditions and resistance to infection.

O presente estudo comparou características hematológicas de juvenis de jundiás Rhamdia quelen diploides e triploides, um importante peixe de água doce cultivado no sul do Brasil. A morfometria hematológica dos eritrócitos foi determinada em extensões sanguíneas sob microscopia óptica comum. O sangue dos animais foi também utilizado para mensurar o número total de eritrócitos em câmera de Neubauer, bem como os números totais de leucócitos e trombócitos pelo método indireto. Os resultados demonstraram que a triploidia aumentou (p < 0,01) o tamanho e volume dos eritrócitos. No entanto, como esperado, a triploidia reduziu (p < 0,01) o número de eritrócitos, leucócitos e trombócitos circulantes do jundiá. Além disto, a contagem diferencial de células sanguíneas foi diferente em diploides e triploides, sugerindo que a triploidia afeta o número das células, bem como sua proporção no sangue. Linfócitos foram as células predominantes na contagem de leucócitos de peixes diploides (62,5 por cento), enquanto os monócitos foram os predominantes nos peixes triploides (49,6 por cento). Estes resultados sugerem desempenhos diferentes entre as ploidias do jundiá e exige estudos futuros para avaliar o potencial dos jundiás triploides sob condições de cultivo e frente à infecção.

Triploidy in the hematology of jundia juveniles (Siluriformes: Heptapteridae)

This study compared the hematological characteristics of diploid and triploid of jundia, Rhamdia quelen juveniles, an important freshwater fish cultured in south Brazil. Hematological morphometry of erythrocytes were determined in blood smears under a light microscope. The blood was used to measure the number of red blood cells (RBC) with a hemocytometer Neubauer chamber, and the numbers of white blood cells (WBC) and thrombocytes that were obtained using an indirect method. The results showed that triploidy increased (p 0.01) the size and volume of the erythrocytes. Nevertheless, as expected, triploidy decreased (p 0.01) the number of circulating erythrocytes, leucocytes and trombocytes in the blood of jundia. Moreover differential cell counts were different in diploids and triploids, suggesting that triploidy affects the number of cells and their proportion in blood. Lymphocytes were the most predominant cells in the differential counting of diploid fish (62.5%) while monocytes were predominant in triploid fish (49.6%). These results suggest performance differences between ploidies of jundia, and require future studies to evaluate the potential of triploid jundia in the culture conditions and resistance to infection.

O presente estudo comparou características hematológicas de juvenis de jundiás Rhamdia quelen diploides e triploides, um importante peixe de água doce cultivado no sul do Brasil. A morfometria hematológica dos eritrócitos foi determinada em extensões sanguíneas sob microscopia óptica comum. O sangue dos animais foi também utilizado para mensurar o número total de eritrócitos em câmera de Neubauer, bem como os números totais de leucócitos e trombócitos pelo método indireto. Os resultados demonstraram que a triploidia aumentou (p 0,01) o tamanho e volume dos eritrócitos. No entanto, como esperado, a triploidia reduziu (p 0,01) o número de eritrócitos, leucócitos e trombócitos circulantes do jundiá. Além disto, a contagem diferencial de células sanguíneas foi diferente em diploides e triploides, sugerindo que a triploidia afeta o número das células, bem como sua proporção no sangue. Linfócitos foram as células predominantes na contagem de leucócitos de peixes diploides (62,5%), enquanto os monócitos foram os predominantes nos peixes triploides (49,6%). Estes resultados sugerem desempenhos diferentes entre as ploidias do jundiá e exige estudos futuros para avaliar o potencial dos jundiás triploides sob condições de cultivo e frente à infecção.

Natural triploidy in Leporinus cf. elongatus bearing sex chromosomes

Although several cases of natural triploidy in fish have already been described, spontaneous polyploidy in species with differentiated sex chromosomes are rare. We report the occurrence of a triploid fish (3n = 81) Leporinus cf. elongatus, a species characterized by a highly differentiated ZZ/ZW sex chromosome system, from the São Francisco river. The occurrence of a ZZZ triploid adult indicates the viability of this chromosome constitution in this fish.

Sexual polyploidization in red clover

Because sexual polyploidization broadens genetic basis and supply plant breeders with more variability for the selection process, it can be useful in red clover breeding. This paper reports results of three crossing cycles, starting from a parental generation of tetraploid red clover plants (female parent), and diploids from the Quiñiqueli cultivar, selected for production of more than 1% of giant pollen grains (male parent) aiming to obtain tetraploid plants to be used in red clover breeding programs. Crosses in the next generations were performed by mutual cross-pollinations. Chromosome number chimerism and high pollen sterility were detected in F1, F2 and F3, but there was a trend towards increasing seed production and seed viability along the generations, probably due to successful competition between fertile and sterile gametes. The identification of fertile triploids, as well as their recurrent formation along the generations, indicates that triploid block is not complete in red clover, and that triploids may be successfully used as a bridge for the production of sexual polyploids.

Porque a poliploidização sexual amplia a base genética e proporciona aos melhoristas maior variabilidade para o processo de seleção, ela pode ser uma ferramenta útil ao melhoramento de trevo vermelho. Com o objetivo de obter plantas tetraplóides que possam ser utilizadas em programas de melhoramento de trevo vermelho, este trabalho relata resultados de três ciclos de cruzamentos, partindo de uma população parental de plantas tetraplóides de trevo vermelho, como genitores femininos, e de diplóides da cultivar Quiñiqueli, selecionados para produção de mais de 1% de grãos de pólen gigantes, como genitores masculinos. Nas outras gerações, os cruzamentos foram realizados por polinizações cruzadas mútuas. Quimerismo para número cromossômico e alta esterilidade de pólen foram detectados em F1 , F2 e F3, mas houve uma tendência para aumento da produção e viabilidade das sementes ao longo das gerações, provavelmente devido à competição bem sucedida entre gametas férteis e estéreis. A identificação de triplóides férteis, assim como sua formação recorrente ao longo das gerações, indica que o bloco triplóide não é completo em trevo vermelho e que triplóides podem ser utilizados com sucesso para a produção de poliplóides sexuais.

Sexual polyploidization in red clover

Because sexual polyploidization broadens genetic basis and supply plant breeders with more variability for the selection process, it can be useful in red clover breeding. This paper reports results of three crossing cycles, starting from a parental generation of tetraploid red clover plants (female parent), and diploids from the Quiñiqueli cultivar, selected for production of more than 1% of giant pollen grains (male parent) aiming to obtain tetraploid plants to be used in red clover breeding programs. Crosses in the next generations were performed by mutual cross-pollinations. Chromosome number chimerism and high pollen sterility were detected in F1, F2 and F3, but there was a trend towards increasing seed production and seed viability along the generations, probably due to successful competition between fertile and sterile gametes. The identification of fertile triploids, as well as their recurrent formation along the generations, indicates that triploid block is not complete in red clover, and that triploids may be successfully used as a bridge for the production of sexual polyploids.

Porque a poliploidização sexual amplia a base genética e proporciona aos melhoristas maior variabilidade para o processo de seleção, ela pode ser uma ferramenta útil ao melhoramento de trevo vermelho. Com o objetivo de obter plantas tetraplóides que possam ser utilizadas em programas de melhoramento de trevo vermelho, este trabalho relata resultados de três ciclos de cruzamentos, partindo de uma população parental de plantas tetraplóides de trevo vermelho, como genitores femininos, e de diplóides da cultivar Quiñiqueli, selecionados para produção de mais de 1% de grãos de pólen gigantes, como genitores masculinos. Nas outras gerações, os cruzamentos foram realizados por polinizações cruzadas mútuas. Quimerismo para número cromossômico e alta esterilidade de pólen foram detectados em F1 , F2 e F3, mas houve uma tendência para aumento da produção e viabilidade das sementes ao longo das gerações, provavelmente devido à competição bem sucedida entre gametas férteis e estéreis. A identificação de triplóides férteis, assim como sua formação recorrente ao longo das gerações, indica que o bloco triplóide não é completo em trevo vermelho e que triplóides podem ser utilizados com sucesso para a produção de poliplóides sexuais.