Construction of Genetic Linkage Map Based on SSR Markers in Peanut(Arachis hypogaea L.)

Molecular genetic maps of crop species can be used in a variety of ways in breeding and genomic research such as identification and mapping of genes and quantitative trait loci （QTLs） for morphological, physiological and economic traits of crop species. However, a comprehensive genetic linkage map for cultivated peanut has not yet been developed due to the extremely low frequency of DNA polymorphism in cultivated peanut. In this study, 142 recombinant inbred lines （RILs） derived from a cross between Yueyou 13 and Zhenzhuhei were used as mapping population in peanut （Arachis hypogaea L.）. A total 652 pairs of genomic-SSR primer and 392 pairs of EST-SSR primer were used to detect the polymorphisms between the two parents. 141 SSR primer pairs, 127 genomic-SSR and 14 EST-SSR ones, which can be used to detect polymorphisms between the two parents, were selected to analyze the RILs population. Thus, a linkage genetic map which consists of 131 SSR loci in 20 linkage groups, with a coverage of 679 cM and an average of 6.12 cM of inter-maker distance was constructed. The putative functions of 12 EST-SSR markers located on the map were analyzed. Eleven showed homology to gene sequences deposited in GenBank. This is the first report of construction of a comprehensive genetic map with SSR markers in peanut （Arachis hypogaea L.）. The map presented here will provide a genetic framework for mapping the qualitative and quantitative trait in peanut.

Arbuscular mycorrhizal fungi combined with exogenous calcium improves the growth of peanut(Arachis hypogaea L.)seedlings under continuous cropping

The growth and yield of peanut are negatively affected by continuous cropping.Arbuscular mycorrhizal fungi(AMF)and calcium ions(Ca^(2+))have been used to improve stress resistance in other plants,but little is known about their roles in peanut seedling growth under continuous cropping.This study investigated the possible roles of the AMF Glomus mosseae combined with exogenous Ca^(2+)in improving the physiological responses of peanut seedlings under continuous cropping.G.mosseae combined with exogenous Ca^(2+)can enhance plant biomass,Ca^(2+)level,and total chlorophyll content.Under exogenous Ca^(2+)application,the F_v/F_m in arbuscular mycorrhizal(AM)plant leaves was higher than that in the control plants when they were exposed to high irradiance levels.The peroxidase,superoxide dismutase,and catalase activities in AM plant leaves also reached their maximums,and accordingly,the malondialdehyde content was the lowest compared to other treatments.Additionally,root activity,and content of total phenolics and flavonoids were significantly increased in AM plant roots treated by Ca^(2+)compared to either G.mosseae inoculation or Ca^(2+)treatment alone.Transcription levels of AhCaM,AhCDPK,AhRAM1,and AhRAM2 were significantly improved in AM plant roots under exogenous Ca^(2+)treatment.This implied that exogenous Ca^(2+)might be involved in the regulation of G.mosseae colonization of peanut plants,and in turn,AM symbiosis might activate the Ca^(2+)signal transduction pathway.The combination of AMF and Ca^(2+)benefitted plant growth and development under continuous cropping,suggesting that it is a promising method to cope with the stress caused by continuous cropping.

Drought-induced responses of organic osmolytes and proline metabolism during pre-flowering stage in leaves of peanut (Arachis hypogaea L.)

Peanut （Arachis hypogaea L.）, an improtant oil crop, usually encounters drought stress in the process of growth and development, especially at pre-flowering stage. In order to gain insight into the drought tolerance potentials based on osmolyte accumulation and metabolism of proline aspects of peanut, pot experiments were conducted with a split-plot design in Tai＇an, Shangdong Province, China in 2013 and 2014. Pre-flowering drought （PFD） stress and optinum irrigation （control, CK） were served as the main plots and the two peanut cultivars Shanhua 11 and Hua 17 served as sub-plots. Shanhua 11 was drought-tolerant cultivar and Hua 17 was drought-sensitive. The content of soluble sugars, soluble protein, free proline and other free amino acids, the activities of enzymes involved in proline metabolism, and malondialdehyde （MDA） content and ion leakage were all investigated in the two cultivars at pre-flowering stage. Results showed that PFD stress significantly increased the levels of soluble protein, free proline and free amino acid, and increased Al-pyrroline-5-car- boxylate synthetase （P-5-CS, EC 2.7.2.11） activity in the leaves of drought-tolerant and drought-sensitive cultivars. The activity of proline dehydrogenase （proDH） （EC 1.5.99.8） decreased under PFD stress in both cultivars. The leaves of the tolerant cultivar maintained higher increments of osmolyte levels, lower increments of MDA content and ion leakage, as well as a higher increased proportion of P-5-CS activity and higher inhibited proportion of proDH activity under water stress compared with the drought-sensitive cultivar. The study suggests that proline accumulation in peanut leaves under PFD can be explained by the higher enhanced activities of P-5-CS and higher inhibition of proDH. The results will provide useful information for genetic improvement of peanut under drought tolerance.

Functional Analysis of the Phosphoenolpyruvate Carboxylase on the Lipid Accumulation of Peanut(Arachis hypogaea L.) Seeds

Phosphoenolpyruvate carboxylase（PEPC;EC 4.1.1.31） catalyses phosphoenolpyruvate（PEP） to yield oxaloacetate,which is involved in protein biosynthesis.Pyruvate kinase（PK;EC 2.7.1.40） catalyzes PEP to yield pyruvate,which is involved in fatty acid synthesis.In this study,five PEPC genes（AhPEPC1,AhPEPC2,AhPEPC3,AhPEPC4,and AhPEPC5） from peanut have been cloned.Using a quantitative real-time RT-PCR approach,the expression pattern of each gene was monitored during the seed development of four peanut varieties（E11,Hebeigaoyou,Naihan 1,and Huayu 26）.It was found that these five genes shared similar expression behaviors over the developmental stages of E11 with high expression levels at 30 and 40 d after pegging（DAP）;whereas these five genes showed irregular expression patterns during the seed development of Hebeigaoyou.In Naihan 1 and Huayu 26,the expression levels of the five genes remained relatively high in the first stage.The PEPC activity was monitored during the seed development of four peanut varieties and seed oil content was also characterized during whole period of seed development.The PEPC activity followed the oil accumulation pattern during the early stages of development but they showed a significantly negative correlation thereafter.These results suggested that PEPC may play an important role in lipid accumulation during the seed development of four peanut varieties tested.

Sedative effects of peanut (Arachis hypogaea L.) leaf aqueous extracts on brain ATP, AMP, Adenosine and Glutamate/GABA of rats

Peanut (Arachis hypogaea L.) leaf aqueous extracts (PLAE) has been reputed to be a type of sleep-aid in China. To investigate the sedative effects and effect pathways of PLAE, rats (n = 31) were employed in two experiments and intragastrically administrated of (1) distilled water, PLAE (500 mg/kg body weight (BW)) and peanut stem aqueous extracts (PSAE, 500 mg/kg BW);(2) 0, 100 or 500 mg/kg BW of PLAE, respectively for at least 14 days. Six relevant neurotransmitters were measured finally. Experiment-1 (n = 16) results showed that the brain Lactate were significantly elevated (p < 0.05) in rat cerebrums after PLAE administrations, compared with Control and PSAE groups. In respect of brain energy system, significant degradations of the brain adenosine triphos- phate (ATP) (p < 0.05) were observed in the brainstems and even the whole brains of rats though PLAE treatments. Moreover, we found that the brain Adenosine monophosphate (AMP) were clearly decreased (p < 0.05) in rat cerebrum and brainstem regions, while the brain Adenosine revealed an increasing propensity (p = 0.076) in the cerebrums of freely behaving rats. After experiment-2 (n = 15), the γ-aminobutyric acid (GABA) concentrations were statistically (p < 0.05) enhanced and the ratios of Glutamate/GABA were simultaneously reduced (p < 0.05) in rat brainstems, no matter which one dose (100 or 500 mg/kg BW) of PLAE were used. Results indicated that PLAE could influence the target neurotransmitters that related to rat circadian rhythms in the specific brain regions, possessing the potentialities as a sedative or sleep-aid for hypnic therapy purposes.

Identification of main effect and epistatic QTLs controlling initial flowering date in cultivated peanut(Arachis hypogaea L.)

Initial flowering date(IFD)is closely related to mature period of peanut pods.In present study,a population of recombinant inbred lines(RIL)derived from the cross between Silihong(female parent)and Jinonghei 3(male parent)was used to map QTLs associated with IFD.The RIL population and its two parental cultivars were planted in two locations of Hebei Province,China from 2015 to 2018(eight environments).Based on a high-density genetic linkage map(including 2996 SNP and 330 SSR markers)previously constructed in our laboratory,QTLs were analyzed using phenotypic data and the best linear unbiased prediction(BLUP)value of initial flowering date by inclusive composite interval mapping(ICIM)method.Interaction effects between every two QTLs and between individual QTL and environment were also analyzed.In cultivated peanut,IFD was affected by genotypic factor and environments simultaneously,and its broad sense heritability(h2)was estimated as 86.8%。Using the IFD phenotypic data from the eight environments,a total of 19 QTLs for IFD were detected,and the phenotypic variation explained(PVE)by each QTL ranged from 1.15 to 21.82%.Especially,five of them were also detected by the BLUP value of IFD.In addition,12 additive QTLs and 35 pairs of epistatic QTLs(62 loci involved)were identifed by the joint analysis of IFD across eight environments.Three QTLs(qIFDB04.1,qIFDB07.1 and qIFDB08.1)located on chromosome B04,B07 and B08 were identified as main-effect QTL for IFD,which had the most potential to be used in peanut breeding.This study would be helpful for the early-maturity and adaptability breeding in cultivated peanut.

Cloning, Expression Pattern Analysis and Subcellular Localization of Resveratrol Synthase Gene in Peanut (<i>Arachis hypogaea</i>L.)

Resveratrol synthase (RS) is a key enzyme that plays a critical role in the resveratrol synthesis pathway. In this study, six RS genes were isolated and characterized from peanut variety “Zhenzhu Hong” by silico cloning and RT-PCR. Bioinformatics analysis showed that deduced amino acid sequences of the six cloned RS genes were highly conserved with a similarity from 95% to 99% when compared to the RS genes which had been deposited at the GenBank. The results of amino acid sequences analysis showed six RS proteins contained the Chal_Sti_Synt_N and ACP_Syn_III_C domains and can be classified to same family but with different evolutionary distance. Expression pattern analysis by QRT-PCR provided evidence indicating that the mRNA of six RS genes were primarily expressed in the peanut shell at different developmental stages with different expression levels, but only lower levels of them were evident in the peanut kernel. The subcellular localization of RS protein in onion epidermal cell was performed by Agrobacterium tumefaciens-mediated transformation and the green fluorescent was monitored by confocal fluorescence microscopy. The results indicated that, RS1 and RS5 were located in the nucleus and plasma membrane respectively, while the RS2, RS3, RS4 and RS6 were located in both nucleus inner membrane and plasma membrane. The data will provide basic information for elucidating the regulatory mechanisms and enzyme kinetics underlying the RS genes in the resveratrol synthase pathway.

Stress Responses of Peanut (<i>Arachis hypogaea</i>L.) Genotypes as Measured by Trigonelline Content after Exposure to UV-B Radiation

UV-B radiation has been widely documented as a stressor for plants that can cause decreased biomass, reduction in photosynthesis, and oxidative stress. Trigonelline is a secondary metabolite that is biosynthesized in some plants in response to abiotic stress such as UV-B irradiation. The objectives of this study were to examine biochemical stress responses for peanut plants (Arachis hypogaea L.) of four different genotypes (Spanish, Valencia, Virginia, and Runner) after exposure at various lengths to UV-B radiation and to examine the alteration of trigonelline biosynthesis due to the age of the plants. Peanut plants from the genotypes were exposed to UV-B radiation at three exposure times (60, 120, and 180 min);plants from two growth stages, the flowering (R1) and early maturity (R7), were used. Significant positive correlations (rs 0.29-0.74, P≤0.05) were found for trigonelline concentrations and UV-B exposure times. With longer exposure times of 180 min for plants at R7, trigonelline biosynthesis began as early as 10 days after treatment with 154.6 μg·g-1 DW and remained or increased by up to 71.5 μg·g-1 DW (46.3%) throughout the sampling intervals (10, 20, 30, 40, and 50 days after treatment) to a final value of 226.1 μg·g-1 DW. All four genotypes at R7 exhibited trigonelline concentrations 47.3% to 52.4% (71.6 to 96.5 μg·g-1 DW) higher than individuals at R1. Trigonelline biosynthesis at R7 was significantly (P<0.05) affected by all levels of UV-B exposure, whereas trigonelline concentrations at R1 were significantly influenced (P<0.05) by only the longer exposure times (120 and 180 min). No statistically significant difference was found in trigonelline concentration among the four different genotypes. UV-B irradiation had the greatest effect on plants at R7 after 120 and 180 min of exposure, as 15 out of 20 (75%) individuals had significantly higher (P<0.05) trigonelline concentrations.

Determination and Analysis of Contents of Fatty Acids in Peanut(Arachis hypogaea Linn.)by Gas Chromatography

The contents of five kinds of fatty acids(palmitic acid,stearic acid,oleic acid,linoleic acid and linolenic acid)in 56 collected peanut(Arachis hypogaea Linn.)varieties were determined by gas chromatography.The results showed that in the 56 peanut varieties,oleic acid content was in the range of 36.859%-67.093%;linoleic acid content was in the range of 14.122%-61.025%;palmitic acid content was in the range of 8.583%-20.286%;stearic acid content was in the range of 2.442%-8.971%;and linolenic acid content was in the range of 0.028%-0.093%.Peanut samples No.9304,9355 and 9353 had higher oleic acid/linoleic acid(O/L)ratios,which were 4.751,3.623 and 3.049,respectively,while peanut sample No.9337 exhibited the lowest O/L value of 0.899.

Gomparative analysis of DGAT3（diacylglycerol acyltraiisferase 3） gene from different peanut （Arachis hypogaea L.） varieties

Cultivated peanut is one of the primary sources of vegetable oil and protein in developing countries. DG〉A73 family in peanut cotyledons has no membrane-bound regions suggesting that cytosol is one of the sites for triacylglycerol （TAG） biosynthesis in oilseeds. According to functional annotation and classification of 5 cDNA libraries, 12 unigenes were found with relation of peanut DGAT3 in different organs. Three clones of unigenes, OCP- contig168t OCPcontig12101-3 and OCPcontigl2388-1 were selected for sequencing. Full length sequence of DGAT3 was obtained, showing over 98% sequence similarity with peanut DGAT3 gene AY875644 or EU183333. Upon cluster analysis, DGAT3 of 40 culti- vars were divided into 3 types, namely AhDGATS-1, AhDGAT3-2 and AhDGAT3-3. Coding regions are 1023, 1038 and 1026 base pairs which encoded proteins with 340-, 345- and 341-amino acids, respectively. DGAT3-3 might be a novel gene type among the DGAT3 family which provides great help for studying DGAT3 gene evolution in peanut.

不同土壤类型对花生(Arachis hypogaea L.)籽实镉积累特性的影响

为了在不同土壤环境下选育出低镉(Cd)积累型花生品种以提高花生品质,以‘白沙1016号’品种花生(Arachis hypogaea L.)为供试作物,棕壤和潮土为供试土壤,采用盆栽试验的方法,研究了2种类型土壤对花生籽实Cd积累特性的影响。结果表明:2种类型土壤条件下的花生籽实生物量差异显著(P<0.05);在供试土壤Cd处理范围内(≤10.0 mg/kg),籽实生物量均随2种土壤Cd处理浓度增加呈"低促高抑"现象。花生籽实对土壤Cd的生物富集系数均随2种土壤Cd处理增加呈显著降低趋势(P<0.05),且土壤类型间差异不显著。花生籽实Cd含量和Cd生物富集量均随2种土壤Cd处理的增加而显著增加(P<0.05),但两者在棕壤介质中均显著大于潮土(P<0.05)。土壤类型对花生籽实Cd积累差异的影响主要体现在籽实Cd含量和生物富集量方面。而土壤pH、氧化还原电位(Eh)等均是影响籽实对Cd积累差异的重要因素。

Optimized production and properties of thermostable alkaline protease from Bacillus subtilis SHS-04 grown on groundnut (Arachis hypogaea) meal

Production of alkaline protease from Bacillus subtilis SHS-04 was investigated under different fermentation conditions involving low-cost substrates with the aim of optimizing yield of enzyme. Maximum enzyme production (1616.21 U/mL) was achieved using groundnut meal (0.75%) as nitrogen source and 0.5% glucose as carbon source at 48 h cultivation period, pH 9, 45 ° C and 200 rpm. The yield was 348% increase over comparable control samples. The alkaline protease had optimum temperature of 60 ° C and remarkably exhibited 80% relative activity at 70 ° C. It was highly thermostable showing 98.7% residual activity at 60 ° C after 60 minutes of incubation at pH 9.0 and was stable in the presence of organic solvents studied. These properties indicate the viability of the protease for biotechnological and industrial applications. The optimized yield of enzyme achieved in this study establishes groundnut meal as potential low-cost substrate for alkaline protease production by B. subtilis SHS-04.

Crossability between Arachis hypogaea L.and Erectoides species A.sp.9990 GKP

Eighteen cultivars representing five botanical types of Arachis hypogaea L.were se-lected as female parents to cross with A.sp.9990,an Erectoides accession with resistance to mul-tiple diseases and pests.All the crosses failed to produce viable seeds mainly due to embryo abor-tions.Three morphologically different types of embryo abortion were recorded.Marked differ-ences were observed between subspecies,and among botanical types and genotypes in hybrid podproduction and embryo development.In terms of pod production per 100 pollinations,subspeciesfastigiata was superior to subspecies hypogaea,and Valencia and Virginia types performed betterthan Spanish and Peruvian types.Fengqiu Yibazhua and Silihong were among the most acceptablegenotypes by showing higher pod productions and delayed embryo abortions.These results sug-gest that through extensive screenings,particular genotypes with higher crossability or lessincompatibility with A.sp 9990 can be obtained,and the use of such genotypes in

Molecular Cloning,Genomic Organization and Functional Analysis of the Ribosomal Protein S30(RPS30) Gene from Arachis hypogaea

The ribosomal proteins are crucial for the maintenance of ribosomal translational efficiency and fidelity.In the study,we characterized the ribosomal protein S30(RPS30)gene from Arachis hypogaea that has been isolated through Genefishing analysis during defense responses to Ralstonia solanacearum.The cDNA of RPS 30 contained a 189 base pair(bp)open-reading frame encoding 62 amino acids.The genomic DNA consists of 272 bp containing two exons and one 83 bp intron.The RPS 30 mRNA transcript was mainly expressed in roots and leaves.The expression level of the RPS 30 mRNA transcripts was up-regulated sharply 6 h after bacterial challenge and was 12 times greater than that of the control group.The phylogenetic analysis for genes encoding proteins showed that RPS30 were conserved within dicotyledonous and monocotyledonous plants.d S extremely exceeded d N in all branches of the tree(d N/d S<1.0),indicating that functional constraint have acted on RPS 30 throughout evolution.

Genetic Diversity of Peanut (Arachis hypogea L.) Cultivars as Revealed by RAPD Markers

The objectives of this study were to evaluate the genetic diversity of the peanut accessions using Random Amplified Polymorphic DNA (RAPD) molecular marker and to evaluate RAPD markers to be used in peanut as genetic markers and improve such techniques as suitable strategies for peanut germplasm characterization. Twenty peanut accessions were included in this study and were subjected to RAPD molecular markers analysis. Twenty-seven RAPD primers produced 210 amplification products of which 80 (36.4%) were polymorphic. In conclusion, this study reported a successful fingerprinting of peanut accessions using RAPD markers and demonstrated the usefulness of these markers in estimating the extent of genetic variation in peanut germplasm.

Development and validation of simple sequence repeat markers from Arachis hypogaea transcript sequences

Simple sequence repeats(SSRs) are important molecular markers for assessing genetic diversity in Arachis hypogaea L. and many other crops and constructing genetic linkage maps for important agricultural traits. In this study, 29,357 potential SSRs were identified in 22,806 unigenes assembled from A. hypogaea transcript sequences. Of these unigenes, 1883 and 4103 were annotated and assigned in Kyoto Encyclopedia of Genes and Genomes Orthology and Eukaryotic Orthologous Groups databases, respectively. Among the SSR motifs, mono-(19,065; 64.94%) and trinucleotide(5033; 17.14%) repeats were the most common, and the three most dominant motifs were A/T(18,358; 62.54%), AG/CT(2804;9.55%), and AAG/CTT(1396; 4.76%). Polymerase chain reaction(PCR) primer pairs were designed for 4340 novel SSR markers and 210 new SSRs were validated using 24 A. hypogaea varieties. Of the 210, 191(91%) yielded PCR products, with 37(18%) identifying polymorphisms. The 37 polymorphic SSR markers detected 146 alleles(2–10 alleles per locus), and the average polymorphic information content was 0.403(with a range of 0.077 to0.819). The new SSRs enrich the current marker resources for A. hypogaea and may also be useful for genetic diversity analysis, functional genomics research, and molecular breeding.

A New Coumestan from Arachis hypogaea L.

A new coumestan, 3, 9-dihydroxy-4, 8-dimethoxycoumestan, was isolated from Arachis hypogaea L. together with two known compounds: 3, 9-dihydroxy-4-methoxycoumestan and 3, 9-dihydroxy-8-methoxycoumestan. The structure was established by spectroscopic methods.

Biomass Industrial Effluent Effect on Carbohydrates,Aminoacids,Nitrite and Nitrite Enzyme Activities of Arachis hypogaea L.

The objectives of the present investigations are to see the effects of biomass power plant effluent on the carbohydrates, aminoacids, nitrite and nitrite enzyme activitiesand proline of Arachis hypogaea L. var TCGS 320 under controlled pot culture methods. Plants were cultivated with 25, 50, 75 and 100% of the effluent and a control without the effluent. The treatment of the crop with 25% of the effluent has shown stimulatory effect on all the biochemical parameters studied. Carbohydrates, starch, aminoacids, protein, nitrate and nitrite reductase enzymatic activities have increased in 10, 15, 20 DAS （days after sowing）. In 25 and 30 DAS all biochemical parameters have decreased, due to environmental factors （rainfall, temperature, humidity, etc.）. The present investigation clearly indicated that the biomass power plant effluent has stimulatory effect on all the biochemical contents at lower concentration, and at higher concentration they have deleterious effects.

Influence of Fly Ash and Growth Regulator with Soil for Determination of Chlorophyll in <i>Arachis hypogaea</i>L.

The present investigation was conducted to find out the effect of varying levels of fly ash and growth hormones on the determination of chlorophylls. The experiments were conducted in pots during 2009-2010 with Arachis hypogaea L. (groundnut) grown with different levels of fly ash concentration, and soil was used (various combinations) at Guru ghasidas University, Bilaspur (CG.) India. In fresh leaf, chlorophylls content varies in the plain soil from 0.29 to 0.64 mg g-1, which is less for photosynthetic activities. Arachis hypogaea L. showed maximum germination percentage, increasing leaf area, enhancement of root & shoot length, whereas Fly ash, bio fertilizers with growth hormone showed minimum values in all parameters. Results showed that, for combination of A to E, the value of chlorophyll ranged from 0.270 mg g-1 to 0.395 mg g-1, and chlorophyll b ranged from 0.400 mg g-1 to 0.489 mg g-1, whereas fro total chloro- phyll ranged from 0.67 to 0.85 mg g-1. In the present work, chlorophyll a, chlorophyll b & total chlorophyll content in fresh leaf, after 45 days, were recorded as 0.395 mg g-1, 0.489 mg g-1 and 0.851 mg g-1 while in 90 days were recorded as 10.38 mg g-1, 0.48 mg g-1 and 0.86 mg g-1 respectively, in less amount combination of fly ash, soil content with application of growth hormone.

Photoperiod Affects <i>in Vitro</i>Flowering in Wild Peanut <i>Arachis paraguariensis</i>

Arachis paraguariensis, a wild peanut species, is a potential experimental system for studying the molecular mechanisms of flowering in the genus Arachis. The present study was carried out to investigate the effect of photoperiod on in vitro reproductive behavior of five genotypes of A. paraguariensis. Day-lengths of 12, 16 and 24 h were tested to monitor in vitro flowering using growth chambers kept at 26?C ± 1?C and 60% ± 5% relative humidity under an illumination of 40 μmol?m–2?s–1. Flowering percentage of plantlets ranged from 35% to 93%, 20% to 75%, and 5% to 53% for 12, 16 and 24 h day-lengths, respectively. Genotype PI 262842 displayed the highest frequency of flowering under all the day-length treatments but in vitro flower bud initiation was delayed. The highest mean flowering percentage of 65% across all the genotypes for plantlets exposed to 12 h photoperiod is indicative that flowering induction actually occurred. The results presented in this paper provide evidence for photoperiodic flowering response as well as the occurrence of short day-length-enhanced flowering in A. paraguariensis.