Breeding of ornamental orchids with focus on Phalaenopsis: current approaches, tools, and challenges for this century.

Ornamental orchid breeding programs have been conducted to develop commercially valuable cultivars with improved characteristics of commercial interest, such as size, flower color, pattern, shape, and resistance to pathogens. Conventional breeding, including sexual hybridization followed by selection of desirable characteristics in plants, has so far been the main method for ornamental breeding, but other techniques, including mutation induction by polyploidization and gamma irradiation, and biotechnological techniques, such as genetic transformation, have also been studied and used in ornamental breeding programs. Orchids are one of the most commercially important families in floriculture industry, having very particular reproductive biology characteristics and being a well-studied group of ornamentals in terms of genetic improvement. The present review focuses on the conventional and biotechnological techniques and approaches specially employed in breeding Phalaenopsis orchids, the genus with highest worldwide importance as an ornamental orchid, highlighting the main limitations and strengths of the approaches. Furthermore, new opportunities and future prospects for ornamental breeding in the CRISPR/Cas9 genome editing era are also discussed. We conclude that conventional hybridization remains the most used method to obtain new cultivars in orchids. However, the emergence of the first biotechnology-derived cultivars, as well as the new biotechnological tools available, such as CRISPR-Cas9, rekindled the full potential of biotechnology approaches and their importance for improve ornamental orchid breeding programs.

Wavelength and Light Intensity Affect Macro- and Micronutrient Uptake, Stomata Number, and Plant Morphology of Common Bean (Phaseolus vulgaris L.).

It is already known that light quality and intensity have major influences on the growth, etiolation, germination, and morphology of many plant species, but there is limited information about the effect of wavelength and light intensity on nutrient absorption by plants. Therefore, this study was established to evaluate the plant growth, stomata formation, chlorophyll index, and absorption of macro- and micronutrients by common bean plants under six light treatments. The experimental design was completely randomized and consisted of six treatments: strong blue (blue LED at high light intensity); weak blue (blue LED at low light intensity); strong red (red LED at high light intensity); weak red (red LED at low light intensity; pink (combined red + blue LED), and white (combined red + white led). The stomatal density (stomata mm-2); the SPAD index; plant height (cm); root length (cm); plant dry weight (g); root dry weight (g); and the concentrations of N, S, K, Mg, Ca, B, Zn, Mn, and Fe on leaf analysis were influenced by all treatments. We found that plant photomorphogenesis is controlled not only by the wavelength, but also by the light intensity. Etiolation was observed in bean plants under blue light at low intensity, but when the same wavelength had more intensity, the etiolation did not happen, and the plant height was the same as plants under multichromatic lights (pink and white light). The smallest plants showed the largest roots, some of the highest chlorophyll contents, and some of the highest stomatal densities, and consequently, the highest dry weight, under white LED, showing that the multichromatic light at high intensity resulted in better conditions for the plants in carbon fixation. The effect of blue light on plant morphology is intensity-dependent. Plants under multichromatic light tend to have lower concentrations of N, K, Mg, and Cu in their leaves, but the final amount of these nutrients absorbed is higher because of the higher dry weight of these plants. Plants under blue light at high intensity tended to have lower concentrations of N, Cu, B, and Zn when compared to the same wavelength at low intensity, and their dry weight was not different from plants grown under pink light. New studies are needed to understand how and on what occasions intense blue light can replace red light in plant physiology.

Insights into cryopreservation, recovery and genetic stability of medicinal plant tissues.

Several plant secondary metabolites are used in the production of different pharmaceuticals based on their biological activities. The conservation and sustainable use of medicinal plants is important for the industrial production of plant-based medicines. Different cryopreservation methods are used for long-term culture preservation, which allows fast regeneration of the preserved plant material with the maintenance of its primary original traits. These methods could ensure the sustainable indefinite supply of plant tissues for theoretically unlimited periods of time, and have gained considerable attention in recent years. It is important to assess the recovery rate and the genetic stability of the recovered plant tissues after cryopreservation because cryopreservation efficiency differs among plant tissues and species. This review lays particular emphasis on the pharmaceutical applications of plant secondary metabolites that are produced through tissue culture approaches, highlighting the methods used for their cryopreservation, as well as their recovery and genetic stability.

Orchid Micropropagation Using Conventional Semi-Solid and Temporary Immersion Systems: A Review.

Orchids, with their astonishingly stunning flowers, dominate the international floricultural market. They are considered prized assets for commercial applications in pharmaceutical and floricultural industries as they possess high therapeutic properties and superior ornamental values. The alarming depletion of orchid resources due to excessive unregulated commercial collection and mass habitat destruction makes orchid conservation measures an extreme priority. Conventional propagation methods cannot produce adequate number of orchids, which would meet the requirement of these ornamental plants for commercial and conservational purposes. In vitro orchid propagation using semi-solid media offers an outstanding prospect of rapidly producing quality plants on a large scale. However, the semi-solid (SS) system has shortcomings with low multiplication rates and high production costs. Orchid micropropagation using a temporary immersion system (TIS) overcomes the limitations of the SS system by reducing production costs and making scaleup and full automation possible for mass plant production. The current review highlights different aspects of in vitro orchid propagation using SS and TIS and their benefits and drawbacks on rapid plant generation.

Artificial pollination and use of growth regulators in atemoya 'Gefner' fruits

The objective was to study the effectiveness of the growth regulator (ANA + GA3) associated or not to the application of adjuvant and artificial pollination in 'Gefner' atemoya. The experiment was conducted in the experimental orchard at Florida's Tropical Research and Education Center. The experimental design was in a randomized block, with 14 treatments, 10 repetitions and 3 flowers per plot. The highest percentages of fixed fruits were obtained with hand pollination ­ HP and 450 NAA + 1250 GA3 mg L-1 + adjuvant and HP. The use of hand pollination for 'Gefner' atemoya tree proved to be the most efficient method so far. Applying growth regulator without artificial pollination produces parthenocarpic fruits, however with high rate of abortions, and small fruits. Growth regulators together with hand pollination produces small and uneven fruits, and cause reduction in the fruits' titratable acidity. The use of adjuvant caused low fixation and toxicity to fruits, and its use is not recommended.

Development and validation of a standard area diagram set as assessment aid for estimating the severity of bacterial spot on tri-lobed leaves of yellow passion fruit / Desenvolvimento e validação de escala diagramática como auxílio na avaliação da severidade da bacteriose em folhas trilobadas do maracujazeiro azedo

Bacterial spot (Xanthomonas axonopodis pv. passiflorae) significantly reduces yellow passion fruit (Passiflora edulis Sims) yield and longevity. A standard area diagram set (SADs) for severity assessment of bacterial spot on tri-lobed leaves of yellow passion was developed and validated in this study. The SADs consisted of eight severity levels (2; 4; 9; 18; 35; 58; 80; and 94%). For its validation, 20 raters, who initially estimated the disease severity without the aid of the SADs, were divided into four groups (G1 and G3, inexperienced; G2 and G4, experienced). Subsequently, G1 and G2 performed the second evaluation without the SADs, and G3 and G4 completed the second evaluation with the proposed SADs. The accuracy and precision of the assessments were determined by simple linear regression and by the Lin's concordance correlation coefficient (LCCC). The proposed SADs allowed accurate and precise quantification of bacterial spot severity, increasing the agreement between estimated and actual values. Inexperienced raters benefited the most from the use of the SADs. The increase in accuracy and precision in the non-aided groups, when present, was less pronounced than those increments observed in the SADs-aided groups. The LCCC confirmed the increases in accuracy and precision detected by the linear regression analysis.

A bacteriose (Xanthomonas axonopodis pv. passiflorae) reduz significativamente a produção e longevidade do maracujazeiro azedo (Passiflora edulis Sims). Uma escala diagramática para a avaliação da severidade da bacteriose em folhas trilobadas do maracujazeiro azedo foi desenvolvida e validada neste estudo. A escala diagramática apresentou oito níveis de severidade (2; 4; 9; 18; 35; 58; 80 e 94%). Para a sua validação, os 20 avaliadores foram divididos em quatro grupos (G1 e G3, sem experiência; G2 e G4, com experiência), que inicialmente estimaram a severidade da doença sem auxílio da escala. Posteriormente, G1 e G2 fizeram outra avaliação sem escala, e G3 e G4 realizaram a avaliação com a escala proposta. A acurácia e a precisão das estimativas foram determinadas por regressão linear simples e pelo coeficiente de correlação de concordância de Lin (LCCC). A escala diagramática proposta permitiu quantificar a severidade da bacteriose de forma acurada e precisa, aumentando a concordância entre os valores estimados e os reais. Os avaliadores inexperientes foram os mais beneficiados pelo uso da escala. O aumento da acurácia e precisão nos grupos que realizaram dupla avaliação sem escala, quando ocorreu, foi mais discreto que os incrementos observados nos grupos que utilizaram a escala. O LCCC confirmou os incrementos da acurácia e precisão detectados pela análise de regressão linear.

Molecular characterization of yellow passion fruit genotypes with different yield and disease resistance levels / Caracterização molecular de genótipos de maracujazeiro azedo com diferentes níveis de produtividade e resistência a doenças

Brazil is the largest passion fruit producer in the world. However, the yield is still considered low, and the cultivation of unsuitable varieties is one of the factors directly influencing this trait. As a consequence, breeding studies have been developed with the purpose of obtaining genetic materials with high yield, high fruit quality, and disease resistance. The objective of this study was to characterize and quantify the genetic variability in 18 genotypes of yellow passion fruit (Passiflora edulis Sims) with different levels of yield and disease resistance, using RAPD markers. The RAPD markers were obtained from 10 decamer primers and converted into a matrix of binary data. Estimations of the genetic dissimilarities between different genotypes and cluster analysis were performed. A total of 58 markers were generated, 63.80% of which were polymorphic. The genetic distances among genotypes varied from 0.040 to 0.354 and genotypes were subdivided into at least 5 groups of similarity. The dispersion graphs showed a low clustering tendency for yield and resistance to different diseases (septoriosis, anthracnose, scab, bacterial spot, and passion fruit woodiness disease). These results demonstrate a high genetic variability among the evaluated genotypes, which is valuable information when selecting promising materials to be used per se or as parents in genetic breeding programs.

O Brasil é o maior produtor mundial de maracujá. Entretanto, a produtividade ainda é considerada baixa e o cultivo de variedades inadequadas é um dos fatores que influenciam diretamente esta característica. Como consequência, trabalhos de melhoramento genético tem sido desenvolvidos com a finalidade de obter materiais genéticos com alta produtividade, qualidade de frutos e resistência a doenças. Este trabalho objetivou caracterizar e quantificar a variabilidade genética em 18 genótipos de maracujazeiro azedo (Passiflora edulis Sims) com diferentes níveis de produtividade e resistência a doenças, utilizando marcadores moleculares RAPD. Os marcadores RAPD, obtidos por meio de 10 iniciadores decâmeros, foram convertidos em uma matriz de dados binários. Estimativas de dissimilaridades genéticas entre os diferentes genótipos e análises de agrupamento foram realizadas. Um total de 58 marcadores foram gerados, dos quais 63,80% foram polimórficos. As distâncias genéticas entre os genótipos variaram de 0,040 a 0,354 e os genótipos foram subdivididas em pelo menos 5 grupos de similaridade. Os gráficos de dispersão mostraram uma baixa tendência de agrupamento para produtividade e resistência à septoriose, antracnose, verrugose, bacteriose e virose do endurecimento dos frutos. Estes resultados demonstram uma alta variabilidade genética entre os genótipos estudados, que é uma informação valiosa para a seleção de materiais promissores para serem utilizados per se ou como parentais em programas de melhoramento genético.

Reaction of yellow passion fruit to passion fruit woodiness disease and to bacterial spot / Reação de maracujazeiro azedo à virose do endurecimento dos frutos e à bacteriose

The use of resistant varieties is a promising strategy for passion fruit woodiness disease (PWD) (Cowpea aphid-borne mosaic virus - CABMV) and bacterial spot (Xanthomonas axonopodis pv. passiflorae - Xap) control in yellow passion fruit (Passiflora edulis Sims). This study aimed at evaluating the reaction of nine genotypes of yellow passion fruit to both mechanically inoculated CABMV and Xap, under protected cultivation. The experiment was conducted using a randomized block design with subdivided parcels comprised of nine treatments, four repetitions, six replications per genotype, and five evaluations. Disease incidence (% plants infected) and severity (% of total leaf area with necrotic lesions or leaf symptoms) were calculated at 7-day intervals. All genotypes evaluated were classified as moderately susceptible to PWD. MAR20#10, MAR20#41, and Rosa Intenso were classified as moderately resistant to bacterial spot whereas the remaining genotypes were classified as moderately susceptible. Genotypes Rosa Intenso, MAR20#41, MAR20#15, and MSCA stood out for presenting the lowest PWD mean severity scores as well as the greatest numbers of plants presenting resistance to PWD after five evaluations. MAR20#10, MAR20#41, and Rosa Intenso demonstrated the lowest bacterial spot mean severity scores. Among the genotypes selected, Rosa Intenso and MAR20#41 were the most promising genotypes for presenting the lowest severity scores for both PWD and bacterial spot disease.

O uso de variedades resistentes é uma estratégia promissora para o controle da virose do endurecimento dos frutos (VEF) (Cowpea aphid-borne mosaic virus - CABMV) e da bacteriose (Xanthomonas axonopodis pv. passiflorae - Xap) no maracujazeiro amarelo (Passiflora edulis Sims). Este estudo objetivou avaliar a reação de nove genótipos de maracujazeiro amarelo à CABMV e Xap, ambos inoculados mecanicamente, sob cultivo protegido. O experimento foi conduzido em delineamento de blocos casualizados com parcelas subdivididas, composto por nove tratamentos, quatro repetições, seis plantas por genótipo e cinco avaliações. A incidência (% plantas infectadas) e a severidade (% da área foliar total com lesões necróticas ou sintomas foliares) das doenças foram calculadas em intervalos de sete dias. Todos os genótipos avaliados foram classificados como moderadamente suscetíveis à VEF. MAR20#10, MAR20#41 e Rosa Intenso foram classificados como moderadamente resistente à bacteriose enquanto os demais genótipos foram classificados como moderadamente suscetíveis. Os genótipos Rosa Intenso, MAR20#41, MAR20#15 e MSCA se destacaram por apresentarem menores severidades médias da VEF bem como pelo maior número de plantas apresentando resistência à virose após as cinco avaliações. MAR20#10, MAR20#41 e Rosa Intenso demonstraram as menores severidades médias de bacteriose. Entre os genótipos selecionados, Rosa Intenso e MAR20#41 foram os mais promissores por apresentarem os menores valores de severidade para a VEF e para a bacteriose.

Effectiveness of a micronutrient delivery system fertilizer in jatropha plants is related to enhanced photosynthesis, gas exchange and biomass allocation / Eficácia de um sistema de fornecimento de micronutrientes em plantas de jatropha está relacionado com incrementos na fotossíntese, troca gasosa e alocação de biomassa

This study aimed at comparing the growth and physiological changes in Jatropha curcas L. (jatropha or physic nut) young plants fertilized or not with a commercial product based on a micronutrient delivery system (MDS), under different doses of NPK. Measurements of growth, chlorophyll content, and leaf gas exchange were performed in the greenhouse, where plants were arranged in a split-split plot design. Plants were grown for 120 days in 3.9 L pots containing local soil, with or without MDS (main plot), combined with NPK doses (0; 1.8; 4.7 and 7.4 g L-1) in subplots. Dose-response curves showed that most variables were positively responsive to NPK doses in plants growing without MDS, whereas slight responses or even opposite behavior was observed in MDS-fertilized plants. MDS application under low NPK doses resulted in higher biomass allocation in leaves and roots, increases in number of leaves and chlorophyll content, plant height, stem diameter, shoot and root dry weight, stomatal conductance, photosynthesis, leaf transpiration, and water use efficiency, as well as decreases in intercellular CO2 in the leaf and vapor-pressure deficit. The enhanced biomass allocation, photosynthesis and gas exchange in MDS-supplemented plants indicates the relevant role played by this fertilizer in jatropha metabolism, resulting in more vigorous plants.

Este estudo teve como objetivo comparar o crescimento e as mudanças fisiológicas em plantas jovens de Jatropha curcas L. (pinhão manso), fertilizadas ou não, com um produto comercial baseado em um sistema de fornecimento de micronutrientes (MDS), sob diferentes doses de NPK. Medidas de crescimento, teor de clorofila e troca gasosa foliar foram realizados em casa de vegetação, onde as plantas foram arranjadas em um delineamento de blocos casualizados com parcela subdividida. As plantas foram cultivadas por 120 dias em potes de 3,9 L contendo solo local, com ou sem MDS (parcela principal), combinado com doses de NPK (0; 1,8; 4,7 e 7,4 g L-1) nas subparcelas. Curvas doseresposta mostraram que a maioria das variáveis responderam positivamente às doses de NPK em plantas crescendo sem MDS, enquanto respostas fracas ou mesmo comportamento oposto foi observado em plantas fertilizadas com MDS. A aplicação de MDS em doses baixas de NPK resultaram em maior alocação de biomassa nas folhas e raízes, aumento no número de folhas e teor de clorofila, altura da planta, diâmetro do caule, pesos secos da parte aérea e raiz, condutância estomatal, fotossíntese, transpiração foliar e eficiência no uso da água, bem como na redução do CO2 intercelular na folha e déficit de pressão de vapor. O aumento na alocação de biomassa, fotossíntese e troca gasosa em plantas suplementadas com MDS indica o papel relevante deste fertilizante no metabolismo de pinhão manso, resultando em plantas mais vigorosas.

Quantitative analysis of jatropha growth: micronutrient delivery system and NPK combined effects / Análise quantitativa de crescimento do pinhão manso: efeitos combinados de um sistema de fornecimento de micronutrientes e NPK

Studies approaching jatropha (Jatropha curcas L.) growth through quantitative analysis parameters are limited, especially regarding the response to different fertilizer types and doses. In order to investigate the effects of a micronutrient delivery system (MDS) fertilizer, a full quantitative analysis of growth in jatropha young plants was performed, comparing this system effectiveness under different NPK doses. Plants were grown in 3.9 L pots containing local soil, with or without MDS (main plot), combined with NPK doses (0; 1.8; 4.7 and 7.4 g L-1) in subplots. Dose-response curves of quantitative analysis variables were generated for three periods of time (40, 80 and 120 days after sown) as a sub-subplot. Quantitative analysis of growth showed that most parameters evaluated in this study were improved by MDS application, resulting in benefits for jatropha initial development, regardless of NPK doses. Even without NPK supplementation or under the lowest dose evaluated (1.8 g L-1), MDS provided better growth of J. curcas plants, being usually equivalent to the highest doses of NPK (4.7 and 7.4 g L-1) without MDS. The effective response of jatropha young plants to MDS supplementation indicates that this kind of fertilizer played a relevant role in the species metabolism, resulting in faster growth and enhanced biomass allocation.

Estudos abordando o desenvolvimento do pinhão manso (Jatropha curcas L.) através de análises de parâmetros quantitativos são limitados, especialmente em relação à resposta a diferentes tipos e doses de fertilizantes. Para investigar os efeitos de um sistema de fornecimento de micronutrientes (MDS), realizou-se uma análise quantitativa completa do crescimento de plantas jovens de pinhão manso comparando a eficácia deste sistema sob diferentes doses de NPK. As plantas foram cultivadas por 120 dias em potes de 3,9 L contendo solo local, com ou sem MDS (parcela principal), combinado com doses de NPK (0; 1,8; 4,7 e 7,4 g L-1) nas subparcelas. Curvas dose-resposta das análises quantitativas das variáveis foram geradas para três períodos (40, 80 e 120 dias após semeadura), como uma subsubparcela. As análises quantitativas de crescimento mostraram que a maioria dos parâmetros avaliados neste estudo foi melhorada pela aplicação do MDS, resultando em benefícios para o crescimento inicial do pinhão-manso, independemente da dose de NPK. Mesmo sem suplementação de NPK ou sob a dose mais baixa avaliada (1,8 g L-1), o MDS proporcionou melhor crescimento do pinhão manso, sendo geralmente equivalente às maiores doses de NPK (4,7 e 7,4 g L-1) sem MDS. A resposta efetiva das plantas jovens do pinhão manso à suplementação com MDS indica que este tipo de fertilizante desempenhou um papel relevante no metabolismo desta espécie, resultando em um crescimento mais rápido e melhor alocação de biomassa.

Cryopreservation of banana's cv Grand Naine in vitro rhizomes

ABSTRACT The preservation of banana genetic material is usually performed through seedlings. However, most banana cultivars do not produce seed and are propagated vegetatively. Therefore, cryopreservation is a feasible technique that allows the preservation of banana genotypes indefinitely. For the success of cryopreservation protocols, the selection of cryoprotectants and pre-freezing techniques are important factor. Therefore, the objective of this study was to verify the effects of different cryoprotectants with and without 1% phloroglucinol and pre-cooling periods on the development of a protocol for cryopreservation of in vitro rhizomes ofMusa accuminata(AAA) cv Grand Naine banana. The addition of 1% phloroglucinol to the cryoprotective solutions, such as PVS2 enhanced recovery of cryopreserved banana rhizomes. In addition, pre-cooling of explants in ice for 3 hours in PVS2 + 1% of phloroglucinol allowed efficient cryopreservation of banana rhizomes, followed by successful recovery and regeneration of in vitro shoots of banana cv Grand Naine.

Cryopreservation of banana's cv Grand Naine in vitro rhizomes.

The preservation of banana genetic material is usually performed through seedlings. However, most banana cultivars do not produce seed and are propagated vegetatively. Therefore, cryopreservation is a feasible technique that allows the preservation of banana genotypes indefinitely. For the success of cryopreservation protocols, the selection of cryoprotectants and pre-freezing techniques are important factor. Therefore, the objective of this study was to verify the effects of different cryoprotectants with and without 1% phloroglucinol and pre-cooling periods on the development of a protocol for cryopreservation of in vitro rhizomes ofMusa accuminata(AAA) cv Grand Naine banana. The addition of 1% phloroglucinol to the cryoprotective solutions, such as PVS2 enhanced recovery of cryopreserved banana rhizomes. In addition, pre-cooling of explants in ice for 3 hours in PVS2 + 1% of phloroglucinol allowed efficient cryopreservation of banana rhizomes, followed by successful recovery and regeneration of in vitro shoots of banana cv Grand Naine.

Enriching Genomic Resources and Marker Development from Transcript Sequences of Jatropha curcas for Microgravity Studies.

Jatropha (Jatropha curcas L.) is an economically important species with a great potential for biodiesel production. To enrich the jatropha genomic databases and resources for microgravity studies, we sequenced and annotated the transcriptome of jatropha and developed SSR and SNP markers from the transcriptome sequences. In total 1,714,433 raw reads with an average length of 441.2 nucleotides were generated. De novo assembling and clustering resulted in 115,611 uniquely assembled sequences (UASs) including 21,418 full-length cDNAs and 23,264 new jatropha transcript sequences. The whole set of UASs were fully annotated, out of which 59,903 (51.81%) were assigned with gene ontology (GO) term, 12,584 (10.88%) had orthologs in Eukaryotic Orthologous Groups (KOG), and 8,822 (7.63%) were mapped to 317 pathways in six different categories in Kyoto Encyclopedia of Genes and Genome (KEGG) database, and it contained 3,588 putative transcription factors. From the UASs, 9,798 SSRs were discovered with AG/CT as the most frequent (45.8%) SSR motif type. Further 38,693 SNPs were detected and 7,584 remained after filtering. This UAS set has enriched the current jatropha genomic databases and provided a large number of genetic markers, which can facilitate jatropha genetic improvement and many other genetic and biological studies.

Genetic structure from the oldest Jatropha germplasm bank of Brazil and contribution for the genetic improvement.

Jatropha is a potential oilseed crop, which requires mitigating factors such as the low genetic variability of the species. The solution runs through the research of Brazilian germplasm. Attention should be given to the germplasm of jatropha the north of Minas Gerais, because this is the oldest national collection and because this region may be a regions of jatropha diversity due to selection pressure arising from environmental adversities. Therefore, the objective of this study was to investigate the genetic diversity of 48 accessions of collection from Empresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG), using SSR and ISSR markers. The results showed low genetic diversity, but some individuals stood out as J. mollissima (48), J. podagrica (47), Mexican accessions (42, 43, 44 and 45) and some national accessions (28, 29, 41 and 46). Therefore, aiming to increase the genetic variability and improve the effectiveness of jatropha breeding programs, it is suggested to explore such as parental accessions to generate commercial hybrids. This fact implies the possibility to support future production of jatropha, since this culture may be an important source of income, especially for small farmers living in semiarid regions of Brazil.

Branching, flowering and fruiting of Jatropha curcas treated with ethephon or benzyladenine and gibberellins.

Jatropha curcas L. has been identified for biofuel production but it presents limited commercial yields due to limited branching and a lack of yield uniformity. The objective of this study was to evaluate the effects of single application of ethephon or a combination of 6-benzyladenine (BA) with gibberellic acid isomers A4 and A7 (GA4+7) on branch induction, flowering and fruit production in jatropha plants with and without leaves. Plants with and without leaves showed differences for growth and reproductive variables. For all variables except inflorescence set, there were no significant statistical interactions between the presence of leaves and plant growth regulators concentration. The total number of flowers per inflorescence was reduced as ethephon concentration was increased. As BA + GA4 +7 concentration increased, seed dry weight increased. Thus, ethephon and BA + GA4 +7 applications appeared to affect flowering and seed production to a greater extent than branching. The inability to discern significant treatment effects for most variables might have been due to the large variability within plant populations studied and thus resulting in an insufficient sample size. Therefore, data collected from this study were used for statistical estimations of sample sizes to provide a reference for future studies.

In vitro conservation of Dendrobium germplasm.

Dendrobium is a large genus in the family Orchidaceae that exhibits vast diversity in floral characteristics, which is of considerable importance to orchid breeders, biotechnologists and collectors. Native species have high value as a result of their medicinal properties, while their hybrids are important as ornamental commodities, either as cut flowers or potted plants and are thus veritable industrial crops. Thus, preservation of Dendrobium germplasm is valuable for species conservation, breeding programs and the floriculture industry. Cryopreservation represents the only safe, efficient and cost-effective long-term storage option to facilitate the conservation of genetic resources of plant species. This review highlights 16 years of literature related to the preservation of Dendrobium germplasm and comprises the most comprehensive assessment of thorough studies performed to date, which shows reliable and reproducible results. Air-drying, encapsulation-dehydration, encapsulation-vitrification, vitrification and droplet-vitrification are the current cryopreservation methodologies that have been used to cryopreserve Dendrobium germplasm. Mature seeds, pollen, protoplasts, shoot primordia, protocorms and somatic embryos or protocorm-like bodies (PLBs) have been cryopreserved with different levels of success. Encapsulation-vitrification and encapsulation-dehydration are the most used protocol, while PLBs represent the main explant explored.

Seedling development and evaluation of genetic stability of cryopreserved Dendrobium hybrid mature seeds.

Vitrification, a simple, fast, and recommended cryopreservation method for orchid germplasm conservation, was evaluated for Dendrobium hybrid "Dong Yai" mature seeds. The genetic stability of regenerated seedlings was also evaluated using flow cytometry. Mature seeds from this hybrid were submitted to plant vitrification solution (PVS2) for 0, 0.5, 1, 2, 3, 4, 5, or 6 h at 0 °C. Subsequently, they were plunged into liquid nitrogen (LN) at -196 °C for 1 h and recovered in half-strength Murashige and Skoog culture medium (1/2 MS), and seed germination was evaluated after 30 days. Seeds directly submitted to LN did not germinate after cryopreservation. Seeds treated with PVS2 between 1 and 3 h presented the best germination (between 51 and 58%), although longer exposure to PVS2 returned moderated germination (39%). Germinated seeds were further subcultured in P-723 culture medium and developed whole seedlings in vitro after 180 days, with no abnormal characteristics, diseases, or nutritional deficiencies. Seedlings were successfully acclimatized under greenhouse conditions with over 80% survival. Flow cytometry analysis revealed no chromosomal changes on vitrified seedlings, as well as seedlings germinated from the control treatment (direct exposure to LN). These findings indicate that vitrification is a feasible and safe germplasm cryopreservation method for commercial Dendrobium orchid hybrid conservation.

Germination, carbohydrate composition and vigor of cryopreserved Caesalpinia echinata seeds

The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood) seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18º or -196ºC and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

Asymbiotic germination response to photoperiod and nutritional media in six populations of Calopogon tuberosus var. tuberosus (Orchidaceae): evidence for ecotypic differentiation.

BACKGROUND AND AIMS: Ecotypic differentiation has been explored in numerous plant species, but has been largely ignored in the Orchidaceae. Applying a specific germination protocol for widespread seed sources may be unreliable due to inherent physiological or genetic differences in localized populations. It is crucial to determine whether ecotypic differentiation exists for restoration and conservation programmes. Calopogon tuberosus var. tuberosus, a widespread terrestrial orchid of eastern North America, is a model species to explore ecotypic differences in germination requirements, as this species occupies diverse habitats spanning a wide geographical range. METHODS: Mature seeds were collected from south Florida, north central Florida, three locations in South Carolina, and the upper Michigan peninsula. Effects of three photoperiods (8/16, 12/12, 16/8 h L/D) were examined on asymbiotic in vitro seed germination and seedling development of C. tuberosus. Germination and early development was monitored for 8 weeks, while advanced development was monitored for an additional 8 weeks. In an additional experiment, asymbiotic seed germination and development was monitored for 8 weeks on six culture media (BM-1 terrestrial orchid medium, Knudson C, Malmgrem, half-strength MS, P723, and Vacin and Went). A tetrazolium test for embryo viability was performed. KEY RESULTS: Short days promoted the highest germination among Florida populations, but few differences among photoperiods in other seed sources existed. Different media had little effect on the germination of Michigan and Florida populations, but germination of South Carolina seeds was higher on media with higher calcium and magnesium. Tetrazolium testing confirmed that South Carolina seeds exhibited low viability while viability was higher in Florida seeds. Seed germination and corm formation was rapid in Michigan seeds across all treatments. Michigan seedlings allocated more biomass to corms compared with other seed sources. CONCLUSIONS: Rapid germination and corm formation may be a survival mechanism in response to a compressed growing season in northern populations. Ecotypic differentiation may be occurring based on seed germination and corm formation data.