Optimizing Nitrogen Application for Jojoba under Intensive Cultivation.

Although jojoba (Simmondsia chinensis) has been cultivated for years, information on its N requirements is limited. A 6-year study of mature jojoba plants grown under field conditions with an intensive management regime evaluated the effect of N application rate on plant nutrient status, growth, and productivity, and nitrate accumulation in the soil. Five levels of N application were tested: 50, 150, 250, 370, and 500 kg N ha-1. Fertilizers were provided throughout the growing season via a subsurface drip irrigation system. Leaf N concentration, in both spring and summer, reflected the level of N applied. A diagnostic leaf (youngest leaf that has reached full size) concentration of 1.3% N was identified as the threshold for N deficiency. Increasing rates of N application resulted in higher P levels in young leaves. Plant K status, as reflected in the leaf analysis, was not affected by N treatment but was strongly affected by fruit load. Vegetative growth was inhibited when only 50 kg N ha-1 was applied. Soil analysis at the end of the fertilization season showed substantial accumulation of nitrate for the two highest application rates. Considering productivity, N costs, and environmental risk, 150 kg N ha-1 is the recommended dosage for intensively grown jojoba. N deficiencies can be identified using leaf analysis, and excess N can be detected via soil sampling toward the end of the growing season. These results and tools will facilitate precise N fertilization in intensive jojoba plantations.

Preventing self-fertilization: Insights from Ziziphus species.

The fitness of self-progeny individuals is inferior to that of their outcrossed counterparts, resulting in a reduction in a plant population's ability to survive and reproduce. To prevent self-fertilization, angiosperms with hermaphrodite flowers may exploit a variety of mechanisms, including synchronous dichogamy and self-incompatibility. Synchronous dichogamy involves two flowering morphs, with strict within-morph synchronization, thereby preventing not only autogamy and geitonogamy but also intra-morph mating. Self-fertilization is also prevented by self-incompatibility, a genetic mechanism that allows the identification and rejection of "self" pollen, thereby preventing both autogamy and geitonogamy. Here, I seek to provide a perspective of flowering in Ziziphus species exhibiting both synchronous (i.e., "Early" morph flowers open in the morning and "Late" morph flowers open in the afternoon) protandrous dichogamy (i.e., pollen dispersal before the stigma becomes receptive) and self-incompatibility.

Impairment of root auxin-cytokinins homeostasis induces collapse of incompatible melon grafts during fruit ripening.

The factors underlying the plant collapse of certain melon-pumpkin graft combinations are not fully understood. Our working hypothesis was that impairment of photoassimilates transport in incompatible combinations induces an imbalance in the homeostasis of root auxin (indole-3-acetic acid; IAA) and of cytokinins, probably triggering plant collapse. Root IAA and cytokinins levels in the presence and absence of fruit and changes in root and scion metabolites were investigated in compatible and incompatible combinations. We showed elevated levels of IAA, 2-oxoindole-3-acetic acid (IAA catabolite), indole-3-acetylaspartate (IAA conjugate), and cis-zeatin-type cytokinins, but low levels of trans-zeatin-type cytokinins in the roots of plants of the incompatible combination during fruit ripening. Similarly, during fruit ripening, the expression of the YUCCA genes, YUC2, YUC6, and YUC11 (required for auxin biosynthesis), the GRETCHEN-HAGEN3 gene (required for auxin conjugation), and the cytokinin oxidase/dehydrogenase 7 (CKX7) gene (regulates the irreversible degradation of cytokinin) was enhanced in the roots of plants of the incompatible combination. Moreover, in the incompatible combination the fruiting process restricted transport of photoassimilates to the rootstock and induces their accumulation in the scion. In addition, high levels of hydrogen peroxide and malondialdehyde and reduced activity of antioxidant enzymes were observed in the roots of the incompatible graft. Our results showed that the collapse of the incompatible graft combination during fruit ripening is closely associated with a dramatic accumulation of IAA in the roots, which probably elicits oxidative damage and disturbs the balance of IAA and cytokinins that is of critical importance in melon-pumpkin graft compatibility.

Breeding an underutilized fruit crop: a long-term program for Hylocereus.

This review describes three decades of introduction, agro-technology development, breeding and selection of Hylocereus species, known as pitaya or dragon fruit, as an example of a holistic program aimed to develop the horticultural potential of a perennial underutilized fruit crop. Interspecific homoploid and interploid crosses and embryo rescue procedures produced improved hybrids, some of which have been released to farmers. Molecular tools and morphological and phenological comparisons between the parental species and the resulting hybrids provided valuable information on dominant/recessive traits and on genetic relationships that could be exploited for further hybridizations. In addition, Hylocereus were crossed with species of the closely related genus Selenicereus, producing valuable intergeneric hybrids. In situ chromosome doubling resulted in the production of autopolyploid lines, from which an understanding of the effect of increased ploidy on fruit traits and metabolomic profiles was obtained. Gamete-derived lines were produced, adding to our biobank homozygote lines that were subsequently used for further hybridization. Spontaneous chromosome doubling occurred in haploid gamete-derived Hylocereus monacanthus lines and in interspecific interploid Hylocereus megalanthus × H. undatus hybrids obtained from an embryo rescue procedure, resulting in plants with double the expected ploidy. Challenging technical problems were addressed by the development of protocols for DNA isolation, flow cytometry, in situ chromosome doubling, androgenesis, gynogenesis and embryo rescue following interspecific and interploidy crosses. Current research leading to the development of genomics and molecular tools, including a draft genome of H. undatus, is also presented. Perspectives for further development of Hylocereus species and hybrids are discussed.

Rootstock identity in melon-pumpkin graft combinations determines fruit metabolite profile.

Grafting has the potential to improve melon fruit yield and quality, but it is currently held that a lack of compatibility between the rootstock and scion compromises such an effect. To throw light on this subject, we studied melon-pumpkin graft combinations with different levels of compatibility to assess to the effect of the rootstock identity on melon fruit yield and quality, including total fruit ortho-diphenols, total flavonoids, and primary fruit metabolites. Melon cv. 'Kiran' (Ki) was grafted onto three pumpkin rootstocks, 'TZ-148' (TZ), 'Shimshon' (Sh), and '53006' (r53), characterized by high, moderate, and low compatibility, respectively. The non-grafted melon cultivar Ki was used as the control. The incompatible combination Ki/r53 gave the lowest fruit yield and the lowest average fruit weight. In that combination, the content of total ortho-diphenols increased vs. Ki and Ki/TZ and that of total flavonoids decreased vs. Ki/Sh. In addition, concentrations of the amino acids, glutamate, methionine, valine, alanine, glycine, and serine, increased in the pulp of the two compatible combinations, i.e., Ki/TZ and Ki/Sh, suggesting that rootstock identity and compatibility with melon Ki scion modulated amino acid synthesis. Our results show an association between rootstock identity (and level of compatibility with the scion) and an enhancement of fruit nutritional values, i.e., high concentrations of organic acids (determined as citrate, malate, fumarate, and succinate) and soluble carbohydrates (sucrose) were recorded in the pulp of the two compatible combinations, i.e., Ki/TZ and Ki/Sh.

The metabolic reserves, carbohydrate balance and nutritional status of jojoba (Simmondsia chinensis), in relation to its annual cycle and fruit load.

Jojoba (Simmondsia chinensis (Link) Schneider) holds high industrial value and an extended cultivation trend. Despite its increased importance, there is a lack of fundamental information about its metabolic reserves and development. Our objective was to characterise metabolite allocation and fluctuations in the carbohydrate and nutrient balance of jojoba plants, as affected by fruit load and the plant's annual cycle. Metabolite profiles were performed for each organ. Soluble carbohydrates (SC) and starch concentrations were surveyed in underground and aboveground organs of high-yield and fruit-removed plants. Simultaneously, nitrogen, potassium and phosphorus were determined in the leaves to evaluate the plant's nutritional status. We found that sucrose and pinitol were the most abundant sugars in all jojoba organs. Each sugar had a 'preferred' organ: glucose was accumulated mainly in the leaves, sucrose and pinitol in woody branches, and fructose in the trunk wood. We found that fruit load significantly influenced the carbohydrate levels in green branches, trunk wood and thin roots. The phenological stage strongly affected the SC-starch balance. Among the examined minerals, only the leaf potassium level was significantly influenced by fruit load. We conclude that jojoba's nutrient and carbohydrate balance is affected by fruit load and the phenological stage, and describe the organ-specific metabolic reserves.

Does scion-rootstock compatibility modulate photoassimilate and hormone trafficking through the graft junction in melon-pumpkin graft combinations?

The effect of the rootstock on the acropetal and basipetal transport of photoassimilates and hormones was studied in the 'Kiran' (Ki) melon cultivar grafted onto pumpkin rootstocks with different degrees of compatibility. A complementary experiment was performed to compare the incompatible combination (as evidenced by plant collapse at the fruit ripening stage), designated Ki/r53, with self-grafted r53/r53 as a model compatible combination. Both experiments showed the accumulation of a number of amino acids, sugars, and sugar alcohols in the scion of the incompatible Ki/r53 grafts. Additionally, they showed a marked reduction of trans-zeatin-type cytokinins and an elevated content of cis-zeatin-type cytokinins in the rootstock, and the opposite pattern in the scion, hinting at the possible involvement of a hormonal signal for graft compatibility. There was no direct evidence of a blockage at the graft union, since hormone acropetal and basipetal trafficking was demonstrated for all combinations. Dye uptake experiments did not show xylem flow impairment. A possibly significant finding in the incompatible combination was the deposition of undifferentiated cells in the hollow space that replaces the pith region in melon and pumpkin. The link between the above findings and the collapse of the plants of the incompatible combination remains unclear.

Grafting in Hylocereus (Cactaceae) as a tool for strengthening tolerance to high temperature stress.

The Hylocereus species that are grown as exotic fruit crops are very often farmed under marginal agronomic conditions, which may include exposure to high temperatures. Here we present a pioneering investigation of grafting as an agro-technique to improve heat tolerance in Hylocereus. To this end, we studied the diploid species H. undatus, the tetraploid H. megalanthus and its di-haploid gamete-derived line 2719, and the interspecific-interploid tetraploid Z-10, all grafted onto H. undatus as the rootstock. Self-grafted, grafted and non-grafted plants were acclimated for one week (to obtain baseline values) and then exposed to heat stress (45/35 °C day/night) for three days, followed by a one-week recovery period under optimal temperatures (30/22 °C). A comparison of the physiological, biochemical and molecular performances of the grafted and self-grafted plants under heat stress and during the recovery period vs those of non-stressed plants (control; 30/22 °C) showed that the grafted and self-grafted plants performed better in most of the assessments: grafted and self-grafted plants recovered more rapidly from the heat stress and suffered far less stem damage. An unexpected - but important - finding that may have implications for other crop was that the self-grafted plants showed better performance than non-grafted plants throughout the trial. Our findings provide support for grafting as a strategy for coping with the stress induced by extremely high temperatures. This study thus paves the way for further investigations of grafting in Hylocereus as a valuable technique that will maintain crop productivity in the face of increasing worldwide temperatures.

Intraspecific Seasonal Variation of Flowering Synchronization in a Heterodichogamous Tree.

Heterodichogamous reproduction in plants involves two flowering morphs, reciprocal in their timing of male and female sexual functions. The degree of synchrony in floral sex phase, within and between individuals of each morph, determines the flowers' potential fertilization partners. Complete within-morph synchrony enables across-morph mating alone, whereas unsynchronized floral sex phases may allow fertilization within a plant individual (geitonogamy) or within a morph. We documented the disruption of flowering synchrony in the heterodichogamous Ziziphus spina-christi towards the end of its seven-month flowering season. This desert tree has self-incompatible, protandrous, short-lived (2-day) flowers that open before dawn ('Early' morph) or around noon ('Late' morph). We counted flowers in the male and female phase on flowering branches that were sampled monthly during the 2016-2018 flowering seasons. In 2018, we also tagged flowers and followed their sex-phase distributions over two days at the start, middle, and end of the season. The switch to the female phase was delayed at the end-season (November-December), and 74% of the flowers did not develop beyond their male phase. Differences in male-phase duration resulted in asynchrony among flowers within each tree and among trees of both flowering morphs. Consequently, fertilization between trees of the same morph becomes potentially possible during the end-season. In controlled hand-pollination assays, some within-morph fertilizations set fruit. The end-season breakdown of synchronous flowering generates variability within morphs and populations. We suggest that this variability may potentially enable new mating combinations in a population and enhance its genetic diversity.

Integrated sRNAome and RNA-Seq analysis reveals miRNA effects on betalain biosynthesis in pitaya.

BACKGROUND: MicroRNAs (miRNAs) and their regulatory functions in anthocyanin, carotenoid, and chlorophyll accumulation have been extensively characterized in many plant species. However, the miRNA regulatory mechanism in betalain biosynthesis remains mostly unknown. RESULTS: In this study, 126 conserved miRNAs and 41 novel miRNAs were first isolated from Hylocereus monacanthus, among which 95 conserved miRNAs belonged to 53 miRNA families. Thirty-four candidate miRNAs related to betalain biosynthesis were differentially expressed. The expression patterns of those differential expressed miRNAs were analyzed in various pitaya tissues by RT-qPCR. A significantly negative correlation was detected between the expression levels of half those miRNAs and corresponding target genes. Target genes of miRNAs i.e. Hmo-miR157b-HmSPL6-like, Hmo-miR160a-Hpcyt P450-like3, Hmo-miR6020-HmCYP71A8-like, Hmo-novel-2-HmCYP83B1-like, Hmo-novel-15-HmTPST-like, Hmo-miR828a-HmTT2-like, Hmo-miR858-HmMYB12-like, Hmo-miR858-HmMYBC1-like and Hmo-miR858-HmMYB2-like were verified by 5'RACE and transient expression system in tobacco. CONCLUSIONS: Hmo-miR157b, Hmo-miR160a, Hmo-miR6020 Hmo-novel-2, Hmo-novel-15, Hmo-miR828a and Hmo-miR858 play important roles in pitaya fruit coloration and betalain accumulation. Our findings provide new insights into the roles of miRNAs and their target genes of regulatory functions involved in betalain biosynthesis of pitaya.

In Support of Winge's Theory of "Hybridization Followed by Chromosome Doubling".

Polyploidy-or chromosome doubling-plays a significant role in plant speciation and evolution. Much of the existing evidence indicates that fusion of unreduced (or 2n) gametes is the major pathway responsible for polyploid formation. In the early 1900s, a theory was put forward that the mechanism of "hybridization followed by chromosome doubling" would enable the survival and development of the hybrid zygote by providing each chromosome with a homolog with which to pair. However, to date there is only scant empirical evidence supporting this theory. In our previous study, interspecific-interploid crosses between the tetraploid Hylocereus megalanthus, as the female parent, and the diploid H. undatus, as the male parent, yielded only allopentaploids, allohexaploids, and 5x-and 6x-aneuploids instead of the expected allotriploids. No viable hybrids were obtained from the reciprocal cross. Since H. undatus underwent normal meiosis with regular pairing in the pollen mother cells and only reduced pollen grains were observed, the allohexaploids obtained supported the concept of "chromosome doubling." In this work, we report ploidy level, fruit morphology, and pollen viability and diameter in a group of putative hybrids obtained from an embryo rescue procedure following controlled H. megalanthus × H. undatus crosses, with the aim to elucidate, for the first time, the timing and developmental stage of the chromosome doubling. As in our previous report, no triploids were obtained, but tetraploids, pentaploids, hexaploids, and 5x- and 6x-aneuploids were found in the regenerated plants. The tetraploids exhibited the morphological features of the maternal parent and could not be considered true hybrids. Based on our previous studies, we can assume that the pentaploids were a result of a fertilization event between one unreduced (2n) female gamete from the tetraploid H. megalanthus and a normal (n) haploid male gamete from H. undatus. All the allohexaploids obtained from the embryo rescue technique where those that regenerated from fertilized ovules 10 days after pollination (at the pro-embryo stage), showing that the chromosome doubling event occurred at a very early development stage, i.e., at the zygote stage or shortly after zygote formation. These allohexaploids thus constitute empirical evidence of "hybridization followed by chromosome doubling."

Performance of Hylocereus (Cactaceae) species and interspecific hybrids under high-temperature stress.

High temperatures limit the successful cultivation of the Hylocereus species on a global basis. We aimed to investigate the degree of heat tolerance in three species, namely, the diploids Hylocereus undatus and H. monacanthus, and the tetraploid H. megalanthus, and nine of their interspecific-interploid hybrids. Rooted cuttings were exposed to heat stress (45/35 °C) or control conditions (25/20 °C) for eight days. Initially, the plants were screened for their tolerance to heat stress and ranked into four heat tolerance categories: good tolerance, moderate tolerance, low tolerance, or sensitive, according to the decrease in the maximum quantum efficiency of photosystem II (Fv/Fm) and visual stem damage. The physiological and biochemical performances of the parental species and of three hybrids representing three different heat-tolerance categories were further analyzed in depth. H. megalanthus (classified as heat sensitive) showed a 65% decrease in Fv/Fm and severe visual stem damage, along with a marked reduction in total chlorophyll content, a large increase in malondialdehyde, and inhibition of catalase activity. H. undatus and H. monacanthus, (classified as low-tolerance species) exhibited slight stem "liquification." The good-tolerance hybrid Z-16 exhibited the best performance under heat stress (21% decrease in Fv/Fm) and the absence of stem damage, coupled with a small decrease in total chlorophyll content, a slight increase in malondialdehyde, high antioxidant activity, and proline accumulation progressing with time. Our findings revealed that most of the hybrids performed better than their parental species, indicating that our breeding programs can provide Hylocereus cultivars suitable for cultivation in heat-challenging regions.

Impact of scion/rootstock reciprocal effects on metabolomics of fruit juice and phloem sap in grafted Citrus reticulata.

BACKGROUND: Rootstock has a significant impact on plant growth and development, including fruit maturation. However, the existence of mutual interaction between scion and rootstock is often neglected. To explore the origin of different fruit quality traits in citrus, we studied the effect of rootstock and the reciprocal interaction between scion and rootstock of nine combinations; three mandarin varieties grafted on three different rootstocks. We analyzed the metabolic profile of juice via gas and liquid chromatography-mass spectrometry (GC-MS and LC-MS, respectively). Additionally, we profiled phloem sap composition in the scion and the rootstock. Quality traits of fruit and their physio-chemical characteristics were also evaluated. RESULTS: For all three cultivars, rootstock was found to affect fruit yield and biochemical fruit quality parameters (sugar and acidity) in interactions with the scions. In mandarin juice, eight of 48 compounds (two primary and six secondary) were related directly to the rootstock, and another seven (one primary and six secondary) were interactively affected by scion and rootstock. In scion and rootstock sap, six and 14 of 53 and 55 primary metabolites, respectively, were directly affected by the rootstock, while 42 and 33 were affected by rootstock interactively with scion, respectively. CONCLUSION: In this work, we show for the first time a reciprocal effect between rootstock and scion. Based on our results, the scion and rootstock interaction might be organ, distance or time dependent.

Pollinator Behavior Drives Sexual Specializations in the Hermaphrodite Flowers of a Heterodichogamous Tree.

Dioecy, the specialization of individuals into either male-only or female-only sexual function, has multiple evolutionary origins in plants. One proposed ancestral mating system is heterodichogamy, two morphs of cross-fertilizing hermaphrodite flowers that differ in their timing of flowering. Previous research suggested that small specializations in these morphs' functional genders could facilitate their evolution into separate sexes. We tested the possible role of pollinators in driving such specializations. Ziziphus spina-christi is an insect-pollinated heterodichogamous tree with self-incompatible flowers and two sympatric flowering morphs. We compared the flower development patterns, floral food rewards, pollinator visits, and fruit production between the two morphs. Male-phase flowers of Z. spina-christi's "Early" and "Late" morphs open before dawn and around noon, respectively, and transition into female-phase 7-8 h later. Flowers of both morphs contain similar nectar and pollen rewards, and receive visits by flies (their ancestral pollinators) at similar rates, mostly during the morning. Consequently, the Early morph functions largely as pollen donor. The Late morph, functioning as female in the morning, produces more fruit. We developed an evolutionary probabilistic model, inspired by Z. spina-christi's reproductive system, to test whether pollinator visit patterns could potentially play a role in an evolutionary transition from heterodichogamy towards dioecy. The model predicts that reproductive incompatibility within flowering morphs promotes their evolution into different sexes. Furthermore, the pollinators' morning activity drives the Early and Late morphs' specialization into male and female functions, respectively. Thus, while not required for transitioning from heterodichogamy to dioecy, pollinator-mediated selection is expected to influence which sexual specialization evolves in each of the flowering morphs.

Identification of reliable reference genes for quantitative real-time PCR normalization in pitaya.

BACKGROUND: A suitable reference gene is an important prerequisite for guarantying accurate and reliable results in quantitative real-time PCR (qRT-PCR) analyses. However, there is no absolute universality in reference genes among different species. It's hard to find an ideal reference gene to fit for different tissues and growth periods. Pitaya (Hylocereus) is commercially produced as a new fruit crop at a large scale in tropical and subtropical regions. To date, there is no report on the identification of the most reliable reference genes for qRT-PCR normalization in pitaya. RESULTS: In this study, six candidate reference genes i.e. Actin(1), GAPDH, UBC(1), UBC(2) EF1-α(1) and histone(1) were selected from thirty-nine typical candidate reference genes to determine the most stable reference genes for qRT-PCR normalization in different tissues, temperature stresses and fruit developmental stages of pitaya. Among the six candidate reference genes, Actin(1) and EF1-α(1) were the most stable gene according to calculations of three statistical methods (GeNorm, NormFinder and BestKeeper) while UBC(1) and UBC(2) showed the lowest expression stability. The six candidate reference genes were further validated by comparing expression profiles of key genes related to betalain biosynthesis at flesh coloration stages of Guanhuahong (Hylocereus monacanthus) and Guanhuabai (H. undatus) pitayas. Actin(1) was recommended the best reference gene for accurate normalization of qRT-PCR data. CONCLUSIONS: In this study, the stability of the selected reference genes for normalizing the qRT-PCR data were identified from pitaya. Actin(1) was the most stably expressed genes in different tissues and fruit developmental stages in pitaya. The present work provides the first data of reference gene identification for pitaya and will facilitate further studies in molecular biology and gene function on Hylocereus and other closely related species.

Metabolomic characterization of pitaya fruit from three red-skinned cultivars with different pulp colors.

Pitaya is a new fruit crop, whose exotically colored fruits have excellent nutritional and antioxidant properties. In this study, the primary metabolite profiles of three pitaya cultivars i.e. 'Guanhuahong' (red peel with red pulp), 'Guanhuabai' (red peel with white pulp) and 'Guanhuahongfen' (red peel with pink pulp) were investigated using GC-MS and Ultraviolet-visible spectroscopy. In the fruit pulp, levels of starch, organic acids, and inositol decreased as the fruit matured. Glucose, fructose, sucrose and sorbitol contents increased gradually during fruit maturation and reached their highest levels in the pulp at the mature stage. Citramalic acid was identified for the first time in the pulp of Hylocereus species. Higher levels of total phenols, flavonoids and antioxidant activities were detected in the peel than in the pulp during fruit maturation of all three cultivars. The finding of higher levels of total phenols and flavonoids in the pitaya peel than in the pulp at the mature stage suggests that pitaya peels are a good source of natural phenols and flavonoids.

Homozygote Depression in Gamete-Derived Dragon-Fruit (Hylocereus) Lines.

Putative gamete-derived progenies from two Hylocereus species, the diploid H. monacanthus and the tetraploid H. megalanthus, were studied with the dual aims to confirm their gamete origin and to evaluate their potential use as genetic resources. An additional goal was to determine the origin (allotetraploid vs. autotetraploid) of H. megalanthus by exploring morphological variations in the di-haploid (2x) H. megalanthus progeny. Gamete origin was proved in all five H. monacanthus lines obtained and in 49 of the 70 H. megalanthus lines by using flow cytometry and simple sequence repeat (SSR) markers. The five double-haploid (2x) H. monacanthus lines showed low vigor and abnormal flower development, with malformed ovules and aborted pollen grains. Only one flower set fruit, giving several viable seeds. For H. megalanthus, both abnormal ovules and defective anthers were observed in the di-haploid (2x) and double di-haploid (4x) lines. Among the 46 di-haploid lines, only 14 set fruit. Another 13 di-haploid lines formed flower buds that abscised before anthesis or soon after pollination. The severe sterility of the double-haploid H. monacanthus and the reduced fertility of all the di-haploid and double di-haploid H. megalanthus lines can be linked to their reduced heterozygosity, which drastically affected the development of normal female and male organs. We thus concluded that chromosome doubling, as occurred spontaneously in the double-haploid H. monacanthus and the double di-haploid H. megalanthus, is not sufficient to restore fertility in Hylocereus. We also observed very low gametoclonal variation among the di-haploid (2x) H. megalanthus lines, a finding that supported an autotetraploid, rather than an allotetraploid, origin of this species. Nonetheless, despite the above-described challenging limitations, these gamete-derived lines are currently being bred as the seed parent, offering unique possibilities for genetic research and additional breeding.

Dormancy release and flowering time in Ziziphus jujuba Mill., a "direct flowering" fruit tree, has a facultative requirement for chilling.

In deciduous fruit trees, the effect of chilling on flowering has mostly been investigated in the "indirect flowering" group, characterized by a period of rest between flower bud formation and blooming. In the present study, we explored the effects of chilling and chilling deprivation on the flowering of Ziziphus jujuba, a temperate deciduous fruit tree belonging to the "direct flowering" group, in which flower bud differentiation, blooming and fruit development occur after dormancy release, during a single growing season. Dormancy release, vegetative growth and flowering time in Z. jujuba cv. Ben-Li were assessed following several treatments of chilling. Chilling treatments quantitatively decreased the timing of vegetative bud dormancy release, thereby accelerating flowering, but had no effect on the time from dormancy release to flowering. Trees grown at a constant temperature of 25°C, without chilling, broke dormancy and flowered, indicating the facultative character of chilling in this species. We measured the expression of Z. jujuba LFY and AP1 homologues (ZjLFY and ZjAP1). Chilling decreased ZjLFY expression in dormant vegetative buds but had no effect on ZjAP1expression, which reached peak expression before dormancy release and at anthesis. In conclusion, chilling is not obligatory for dormancy release of Z. jujuba cv. Ben-Li vegetative buds. However, the exposure to chilling during dormancy does accelerate vegetative bud dormancy release and flowering.

Anastatica hierochuntica, an Arabidopsis Desert Relative, Is Tolerant to Multiple Abiotic Stresses and Exhibits Species-Specific and Common Stress Tolerance Strategies with Its Halophytic Relative, Eutrema (Thellungiella) salsugineum.

The search for novel stress tolerance determinants has led to increasing interest in plants native to extreme environments - so called "extremophytes." One successful strategy has been comparative studies between Arabidopsis thaliana and extremophyte Brassicaceae relatives such as the halophyte Eutrema salsugineum located in areas including cold, salty coastal regions of China. Here, we investigate stress tolerance in the desert species, Anastatica hierochuntica (True Rose of Jericho), a member of the poorly investigated lineage III Brassicaceae. We show that A. hierochuntica has a genome approximately 4.5-fold larger than Arabidopsis, divided into 22 diploid chromosomes, and demonstrate that A. hierochuntica exhibits tolerance to heat, low N and salt stresses that are characteristic of its habitat. Taking salt tolerance as a case study, we show that A. hierochuntica shares common salt tolerance mechanisms with E. salsugineum such as tight control of shoot Na+ accumulation and resilient photochemistry features. Furthermore, metabolic profiling of E. salsugineum and A. hierochuntica shoots demonstrates that the extremophytes exhibit both species-specific and common metabolic strategies to cope with salt stress including constitutive up-regulation (under control and salt stress conditions) of ascorbate and dehydroascorbate, two metabolites involved in ROS scavenging. Accordingly, A. hierochuntica displays tolerance to methyl viologen-induced oxidative stress suggesting that a highly active antioxidant system is essential to cope with multiple abiotic stresses. We suggest that A. hierochuntica presents an excellent extremophyte Arabidopsis relative model system for understanding plant survival in harsh desert conditions.