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The concentration of atmospheric CO 2 and temperature are pivotal components of ecosystem productivity, carbon balance, and food security. In this study, we investigated the impacts of a warmer climate (+2 °C above ambient temperature) and an atmosphere enriched with CO 2 (600 ppm) on gas exchange, antioxidant enzymatic system, growth, nutritive value, and digestibility of a well-watered, managed pasture of Megathyrsus maximus, a tropical C 4 forage grass, under field conditions. Elevated [CO 2 ] (eC) improved photosynthesis and reduced stomatal conductance, resulting in increased water use efficiency and plant C content. Under eC, stem biomass production increased without a corresponding increase in leaf biomass, leading to a smaller leaf/stem ratio. Additionally, eC had negative impacts on forage nutritive value and digestibility. Elevated temperature (eT) increased photosynthetic gains, as well as stem and leaf biomass production. However, it reduced P and K concentration, forage nutritive value, and digestibility. Under the combined conditions of eC and eT (eCeT), eT completely offset the effects of eC on the leaf/stem ratio. However, eT intensified the effects of eC on photosynthesis, leaf C concentration, biomass accumulation, and nutritive value. This resulted in a forage with 12% more acid detergent fiber content and 28% more lignin. Additionally, there was a decrease of 19% in crude protein leading to a 15% decrease in forage digestibility. These changes could potentially affect animal feeding efficiency and feedback climate change, as ruminants may experience an amplification in methane emissions. Our results highlight the critical significance of conducting multifactorial field studies when evaluating plant responses to climate change variables., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The reproductive success of a zoophilous plant species depends on biological interaction with pollinators, which involves both the provision and exploitation of flower resources. Currently, there is little information about how future climate change scenarios will impact interactions between plants and their flower visitors in the tropics. This study analyzes the effects of warming and two soil water conditions on interactions between the tropical forage legume species Stylosanthes capitata and its floral visitors during the flowering period. We used a temperature-free air-controlled enhancement (T-FACE) facility to simulate future warming scenarios by increasing canopy temperature. The tested treatments were: irrigated and ambient canopy temperature (Control); non-irrigated and ambient canopy temperature (wS); irrigated and elevated canopy temperature (eT, +2 °C above ambient canopy temperature); and non-irrigated and elevated canopy temperature (wSeT). The effects of treatments on the time of flower opening and closing, sugar concentration in the nectar, and plant-flower visitor interactions were assessed. In the warmed treatments, S. capitata flower opening occurred ~45 min earlier compared to non-warmed treatments, and flowers remained opened for only ~3 h. Further, the sugar concentration in the nectar from eT was 39% higher than in the Control. The effects of warming on floral biology and flower resource production in S. capitata had an impact on the plant-floral visitor relationships with the bees Apis mellifera and Paratrigona lineata, the most abundant potential pollinating floral visitors, and the butterfly visitor Hemiargus hanno. Additionally, around noon, the interactive and additive effects of the combined wS and eT treatments decreased insect visiting frequency. These results suggest that warming and soil water deficiency could affect flower-visitor interactions and thus the reproductive success of S. capitata in tropical belts., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Tropical plant species are vulnerable to climate change and global warming. Since flowering is a critical factor for plant reproduction and seed-set, warming and elevated atmospheric carbon dioxide concentrations (eCO 2 ) are crucial climate change factors that can affect plant reproductive dynamics and flowering related events in the tropics. Using a combined free-air CO 2 enrichment and a free-air temperature-controlled enhancement system, we investigate how warming (+2 °C above ambient, eT) and elevated [CO 2 ] (~600 ppm, eCO 2 ) affect the phenological pattern, plant-insect interactions, and outcrossing rates in the tropical legume forage species Stylosanthes capitata Vogel (Fabaceae). In comparison to the control, a significantly greater number of flowers (NF) per plot (+62%) were observed in eT. Furthermore, in warmed plots flowers began opening approximately 1 h earlier (~09:05), with a canopy temperature of ~23 °C, than the control (~09:59) and eCO 2 (~09:55) treatments. Flower closure occurred about 3 h later in eT (~11:57) and control (~13:13), with a canopy temperature of ~27 °C. These changes in flower phenology increased the availability of floral resources and attractiveness for pollinators such as Apis mellifera L. and visitors such as Paratrigona lineata L., with significant interactions between eT treatments and insect visitation per hour/day, especially between 09:00-10:40. In comparison to the control, the additive effects of combined eCO 2  + eT enhanced the NF by 137%, while the number of A. mellifera floral visits per plot/week increased by 83% during the period of greatest flower production. Although we found no significant effect of treatments on mating system parameters, the overall mean multilocus outcrossing rate (tm = 0.53 ± 0.03) did confirm that S. capitata has a mixed mating system. The effects of elevated [CO 2 ] and warming on plant-pollinator relationships observed here may have important implications for seed production of tropical forage species in future climate scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Anadenanthera (Fabaceae) is endemic to the Neotropics and consists of two tree species: A. colubrina (Vell.) Brenan and A. peregrina (L.) Speg. This study examined the mating system and contemporary gene flow of A. colubrina (Acol) and A. peregrina (Aper) in a highly fragmented area of the Atlantic Forest to provide valuable information that informs conservation strategies. Reproductive adults from forest remnants [n A. colubrina = 30 (2.7 ha), n A. peregrina = 55 (4.0 ha)] and progeny-arrays (n A. colubrina = 322, n A. peregrina = 300) were genotyped for seven nuclear microsatellite markers. Mating system analyses revealed that A. colubrina is a mixed mating species ( t m = 0.619) while A. peregrina is a predominantly outcrossing species ( t m = 0.905). For both Anadenanthera species, high indices of biparental inbreeding were observed (Acol = 0.159, Aper = 0.216), resulting in low effective pollination neighborhood sizes. Categorical paternity analysis revealed different scales of pollen dispersal distance: the majority of crossings occurring locally (i.e., between nearby trees within the same population), with moderate pollen dispersal coming from outside the forest fragments boundaries (Acol mp = 30%, Aper mp = 35%). Nevertheless, pollen immigration from trees outside the populations for both species suggests that the populations are not reproductively isolated. This study highlights the importance of evaluating both mating system and contemporary gene flow for a better understanding of the biology of Anadenanthera species. This information should be considered to ensure the effective conservation and management practices of these plant species., Competing Interests: Alison G. Nazareno is an Academic Editor for PeerJ., (© 2021 Feres et al.)

Here, we explore the mating pattern and genetic structure of a tropical tree species, Cariniana estrellensis, in a small population in which progeny arrays (n=399), all adults (n=28) and all seedlings (n=39) were genotyped at nine highly informative microsatellite loci. From progeny arrays we were able to identify the source tree for at least 78% of pollination events. The gene immigration rates, mainly attributable to pollen, were high, varying from 23.5 to 53%. Although gene dispersal over long distance was observed, the effective gene dispersal distances within the small population were relatively short, with mean pollination distances varying from 69.9 to 146.9 m, and seed dispersal distances occurring up to a mean of 119.6 m. Mating system analyses showed that C. estrellensis is an allogamous species (tm=0.999), with both biparental inbreeding (tm-ts=-0.016) and selfing rates (s=0.001) that are not significantly different from zero. Even though the population is small, the presence of private alleles in both seedlings and progeny arrays and the elevated rates of gene immigration indicate that the C. estrellensis population is not genetically isolated. However, genetic diversity expressed by allelic richness was significantly lower in postfragmentation life stages. Although there was a loss of genetic diversity, indicating susceptibility of C. estrellensis to habitat fragmentation, no evidence of inbreeding or spatial genetic structure was observed across generations. Overall, C. estrellensis showed some resilience to negative genetic effects of habitat fragmentation, but conservation strategies are needed to preserve the remaining genetic diversity of this population.

Mangabeira (Hancornia speciosa Gomes) is a fruit tree of the Apocynaceae family, which is native to Brazil and is a very important food resource for human populations in its areas of occurrence. Mangabeira fruit is collected as an extractive activity, and no domesticated varieties or breeding programs exist. Due to a reduction in the area of ecosystems where it occurs, mangabeira is threatened by genetic erosion in Brazil. The objective of this study was to characterize and evaluate the genetic diversity of 38 mangabeira individuals collected from natural populations in Pernambuco State using inter-simple sequence repeat (ISSR) molecular markers. The ISSR methodology generated a total of 93 loci; 10 were monomorphic and 83 were polymorphic. The average number of loci per primer was 15.5, ranging from 9 (#UBC 866) to 21 (#UBC 834). The results showed a high level of genetic diversity (0.30), and found that only around 30% of genetic variability is distributed among populations (GST = 0.29, ФST = 0.30), with the remainder (ФCT = 70%) found within each population, as expected for forest outcrossing species. Estimates for historic gene flow (1.18) indicate that there is some isolation of these populations, and some degree of genetic differentiation.

Studies of genetic diversity in plant species present in the remaining fragments of the Atlantic Forest are very important for understanding their resilience to such a degraded ecosystem. We analyzed the genetic diversity of 3 populations of the high-density understory species Metrodorea nigra St. Hill. (Rutaceae) located in forest remnants in the region of Ribeirão Preto, SP, Brazil (M13-Rib, BSQ-Rib, and FAC-Crav), by using simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) molecular markers for conservation purposes. A total of 133 polymorphic loci were observed in 136 inter-simple sequence repeat loci (average of 17 per primer). The Nei genetic diversity (HE) was relatively high considering all populations (0.31). The BSQ-Rib population exhibited the highest value (0.27), followed by the M13-Rib (0.26) and FAC-Crav (0.24) populations. The simple sequence repeat markers analyzed showed a high number of alleles (K = 104), with an average of 14.85 alleles per locus. The average observed heterozygosity was 0.516 and the average expected heterozygosity was 0.771, ranging from 0.688 (FAC-Crav) to 0.765 (BSQ-Rib). The fixation indexes showed positive and significant differences from zero for all sample sets, indicating inbreeding, which may have resulted from the species' mating patterns and the barochoric seed dispersal system of M. nigra. Both markers indicated differentiation among populations, with higher values observed for inter-simple sequence repeat markers. No significant differences between juvenile and adult generations in any of the fragments were observed, indicating the resilience of M. nigra to the effects of fragmentation and reduced habitat.

In this analysis, we attempt to understand how monoecy and dioecy drive spatial genetic structure (SGS) in plant populations. For this purpose, plants of the genus Ficus were used as a comparative model due to their particular characteristics, including high species diversity, variation in life histories, and sexual systems. One of the main issues we assessed is whether dioecious fig tree populations are more spatially genetically structured than monoecious populations. Using the Sp statistic, which allows for quantitative comparisons among different studies, we compared the extent of SGS between monoecious and dioecious Ficus species. To broaden our conclusions we used published data on an additional 27 monoecious and dioecious plant species. Furthermore, genetic diversity analyses were performed for two monoecious Ficus species using 12 microsatellite markers in order to strengthen our conclusions about SGS. Our results show that dioecy, more than monoecy, significantly contributes to SGS in plant populations. On average, the estimate of Sp was six times higher for dioecious Ficus species than monoecious Ficus species and it was two times higher in dioecious than monoecious plant species. Considering these results, we emphasize that the long-distance pollen dispersal mechanism in monoecious Ficus species seems to be the dominant factor in determining weak spatial genetic structure, high levels of genetic diversity, and lack of inbreeding. Although Ficus constitute a model species to study SGS, a more general comparison encompassing a wider range of plants is required in order to better understand how sexual systems affect genetic structure.

Copaifera langsdorffii, locally known as copaíba, is a valuable tropical tree with medicinal properties of its oil. We studied the genetic variation, genetic structure, and the mating system of trees in stands of C. langsdorffii (Leguminosae/Caesalpinioideae) located in an extensive area between the Pardo and Mogi-Guaçu basins in São Paulo State, Brazil, and their offspring, conserved in an ex situ germplasm bank at the University of São Paulo in Ribeirão Preto, SP, Brazil, using six microsatellite loci. Leaves were collected from 80 seed trees and from 259 offspring and their DNA extracted. A total of 140 and 175 alleles were found in the seed trees and their offspring, respectively. Low genetic differentiation was observed between stands, indicating intense gene flow due to efficient pollen dispersion vectors. An estimation of the outcrossing rate showed that these stands are outcrossed (tm = 0.98, P > 0.05). The mean variance of the effective population size of each family in two of the stands was 3.69 and 3.43, while the total effective population size retained in the germplasm bank was between 81 and 96. The paternity correlation was low, ranging from 0.052 to 0.148, demonstrating that the families implanted in this germplasm bank are composed predominantly of half-sibs.

Premise of the Study: We developed and characterized nuclear microsatellite markers for Anadenanthera colubrina, a tropical tree species widely distributed in South America., Methods and Results: Leaf samples of mature A. colubrina trees, popularly called "angico," were collected from an area that is greatly impacted by agricultural practices in the region of Ribeirão Preto in São Paulo State in southeastern Brazil. Twenty simple sequence repeat (SSR) markers were developed, 14 of which had polymorphic loci. A total of 96 alleles were detected with an average of 6.86 alleles per polymorphic locus. The expected heterozygosity, calculated at polymorphic loci, ranged from 0.18 to 0.83. Finally, we demonstrated that 18 loci were cross-amplified in A. peregrina., Conclusions: A total of 14 polymorphic markers suggest a high potential for genetic diversity, gene flow, and mating system analyses in A. colubrina.

Metrodorea nigra (Rutaceae) is an endemic Brazilian tree of great ecological importance, frequently found in the submontane regions of ombrophilous dense and semideciduous forests. This tree is useful for reforesting degraded areas and the wood can be employed in construction. We developed 12 microsatellite markers from a genomic library enriched for GA/CA repeats, for this species. Polymorphisms were assessed in 40 trees of a highly fragmented population found in Cravinhos, State of São Paulo, in southeastern Brazil. Among the 12 loci, 8 were polymorphic and only one had fixed alleles in this population. The number of alleles per locus and expected heterozygosity ranged from 2 to 11 and from 0.190 to 0.889, respectively. These results revealed moderate levels of genetic variation in M. nigra population when compared to other tropical species. Additionally, transferability of the 12 primers was tested in seven other Brazilian Rutaceae tree species (endemics: M. stipularis, Galipea jasminiflora, Esenbeckia leiocarpa and non-endemics: E. febrifuga, E. grandiflora, Balfourodendron riedelianum, Zanthoxylum riedelianum). Transferability ranged among species, but at least 8 loci (~67%) amplified in M. stipularis, demonstrating a high potential for transferring microsatellite markers between species of the same genus in the Rutaceae family.

Premise of the Study: This study was designed to develop and characterize a preliminary set of nuclear microsatellite markers for Aspidosperma polyneuron, an endangered tropical tree species., Methods and Results: Leaf samples were collected from a group of mature A. polyneuron trees in a large area along the Pardo and Mogi-Guaçu river basins in São Paulo State in the southeastern region of Brazil. Twenty-five simple sequence repeat (SSR) markers were developed and 16 of 30 samples had polymorphic loci. The number of alleles per locus ranged from two to 17 (average = 7.06). A high diversity (H(e) = 0.65) and parentage exclusion probability for the first (0.9995) and second (0.9999) parents were measured., Conclusions: These 16 polymorphic markers suggest a high potential for genetic diversity, gene flow, and mating system analyses in A. polyneuron.

Twelve microsatellite loci previously developed in the tropical tree Cariniana estrellensis were genetically characterized in Cariniana legalis. Polymorphisms were assessed in 28 C. legalis individuals found between the Pardo and Mogi-Guaçu River basins in the state of São Paulo, Brazil. Of the 12 loci, 10 were polymorphic and exhibited Mendelian inheritance. The allelic richness at each locus ranged from 2-11, with an average of 7 alleles per locus, and the expected heterozygosity ranged from 0.07-0.88. These loci showed a high probability of paternity exclusion. The characteristics of these heterologous microsatellite markers indicate that they are suitable tools for investigating questions concerning population genetics in C. legalis.

Microsatellite markers were transferred and characterized for two Neotropical fig tree species, Ficus citrifolia and Ficus eximia. Our study demonstrated that microsatellite markers developed from different subgenera of Ficus can be transferred to related species. In the present case, 12 of the 15 primer pairs tested (80%) were successfully transferred to both of the above species. Eleven loci were polymorphic when tested across 60 F. citrifolia and 60 F. eximia individuals. For F. citrifolia, there were 4 to 15 alleles per locus, whereas expected heterozygosities ranged from 0.31 to 0.91. In the case of F. eximia, this was 2 to 12 alleles per locus and expected heterozygosities from 0.42 to 0.87.

Microsatellite loci that were previously developed in the tropical tree Tabebuia aurea were used for the genetic analysis of Tabebuia roseo-alba populations. Nine of 10 simple sequence repeat markers were amplified, and the polymorphism was assessed in 58 individuals sampled from two stands in southeastern Brazil. All loci were polymorphic with Mendelian inheritance. The allele numbers were high, ranging from 5 to 13 in population I and 3 to 7 in population II, with means of 8.9 and 5.5, respectively. We conclude that these markers can be efficiently used for parentage and gene-flow studies., (© 2009 The Authors. Journal compilation © 2009 Blackwell Publishing Ltd.)

ABSTRACT New cultivars of the common bean (Phaseolus vulgaris) with durable resistance to anthracnose can be developed by pyramiding major resistance genes using marker-assisted selection. To this end, it is necessary to identify sources of resistance and molecular markers tightly linked to the resistance genes. The objectives of this work were to study the inheritance of resistance to anthracnose in the cultivar TO (carrying the Co-4 gene), to identify random amplified polymorphic DNA (RAPD) markers linked to Co-4, and to introgress this gene in the cultivar Rudá. Populations F(1), F(2), F(2:3), BC(1)s, and BC(1)r from the cross Rudá x TO were inoculated with race 65 of Colletotrichum lindemuthianum, causal agent of bean anthracnose. The phenotypic ratios (resistant/susceptible) were 3:1 in the F(2) population, 1:1 in the BC(1)s, and 1:0 in the BC(1)r, confirming that resistance to anthracnose in the cultivar TO was monogenic and dominant. Six RAPD markers linked to the Co-4 gene were identified, four in the coupling phase: OPY20(830C) (0.0 centimorgan [cM]), OPC08(900C) (9.7 cM), OPI16(850C) (14.3 cM), and OPJ01(1,380C) (18.1 cM); and two in the repulsion phase: OPB03(1,800T) (3.7 cM) and OPA18(830T) (17.4 cM). OPY20(830C) and OPB03(1,800T), used in association as a codominant pair, allowed the identification of the three genotypic classes with a high degree of confidence. Marker OPY20(830C), which is tightly linked to Co-4, is being used to assist in breeding for resistance to anthracnose.

ABSTRACT Three F(2) populations derived from crosses between the resistant cultivar AB 136 and the susceptible cultivar Michelite (MiA), and one F(2) population derived from a cross between AB 136 and Mexico 222 (MeA), were used to identify markers linked to anthracnose resistance genes present in cultivar AB 136. Primer OPZ04 produced a DNA band (OPZ04(560)) linked in coupling phase to the resistance gene for pathotype 89 (8.5 +/- 0.025 cM) in one population derived from the cross MiA. In the same population, primer OPZ09 produced one band (OPZ09(950)) linked in repulsion phase (20.4 +/- 0.014 cM) to the same resistance gene. The simultaneous use of markers in coupling and in repulsion phases allowed the identification of the three genotypic classes. In the other two populations from cross MiA, OPZ04(560) was linked in coupling phase to resistance genes for pathotypes 73 (2.9 +/- 0.012 cM) and 81 (2.8 +/- 0.017 cM). In population MeA, OPZ04(560) was linked in coupling phase (7.5 +/- 0.033 cM) to resistance to pathotype 64. These data suggest that a single gene or complex locus of linked resistance genes present in cultivar AB 136 confers resistance to all four pathotypes of C. lindemuthianum.

Inheritance of anthracnose resistance of the common bean (Phaseolus vulgaris L.) differential cultivar AB 136 to races 89, 64, and 73 (binary system designation) was studied in crosses with the susceptible differential cultivars Michelite (race 89), Mexico 222 (race 64), and Cornell 49-242 (race 73). In each cross two progenitors, the F 1 , F 2 , and backcross-derived plants were inoculated with the respective race under environmentally controlled greenhouse conditions. The results indicate that single dominant gene(s) control resistance to races 89 and 64, giving a segregation ratio of 3:1 in the F 2 , 1:0 in the backcrosses to AB 136, and 1:1 in the backcross to Michelite (race 89), and to Mexico 222 (race 64). For race 73, the following segregation ratios between resistant and susceptible plants were observed: 13:3 in the F 2 , 1:0 in the backcross to AB 136, and 1:1 in the backcross to Cornell 49-242. Such results suggest that two independent genes may determine resistance of AB 136 to race 73, one dominant (Co-6) and one recessive that is proposed to be assigned co-8. Genotypes Co-6_ or co-8 co-8 would condition resistance, whereas susceptibility would be present in genotypes co-6 co-6 Co-8_. Given the dominant nature of anthracnose resistance genes present in line AB 136 and its resistance to 25 races of Colletotrichum lindemuthianum identified in Brazil by other researchers, we included this cultivar as one of the donor parents in our molecular marker-assisted backcross breeding program, to develop common bean cultivars resistant to anthracnose and adapted to Central Brazil.