Effects of cadmium and zinc interactions on the physiological biochemistry and enrichment characteristics of Iris pseudacorus.

Compound pollution with cadmium (Cd) and zinc (Zn) is common in nature. The effects of compounded Cd and Zn on the growth and development of Iris pseudacorus in the environment and the plant's potential to remediate heavy metals in the environment remain unclear. In this study, the effects of single and combined Cd and Zn stress on I. pseudacorus growth and the enrichment of heavy metals in I. pseudacorus seedlings were investigated. The results showed that under Cd (160 µM) and Zn (800 µM) stress, plant growth was significantly inhibited and photosynthetic performance was affected. Cd+Zn200 (160 µM + 200 µM) reduced the levels of malondialdehyde, hydrogen peroxide, and non-protein thiols by 31.29%, 53.20%, and 13.29%, respectively, in the aboveground tissues compared with levels in the single Cd treatment. However, Cd+Zn800 (160 µM + 800 µM) had no effect. Cd and Zn800 inhibited the absorption of mineral elements, while Zn200 had little effect on plants. Compared with that for Cd treatment alone, Cd + Zn200 and Cd+Zn800 reduced the Cd content in aboveground tissues by 54.15% and 49.92%, respectively, but had no significant effect on Cd in the root system. Zn significantly reduced the Cd content in subcellular components and limited the content and proportion of Cd extracted using water and ethanol. These results suggest that a low supply of Zn reduces Cd accumulation in aboveground tissues by promoting antioxidant substances and heavy metal chelating agents, thus protecting the photosynthetic systems. The addition of Zn also reduced the mobility and bioavailability of Cd to alleviate its toxicity in I. pseudacorus.

Increasing Ca2+ accumulation in salt glands under salt stress increases stronger selective secretion of Na+ in Plumbago auriculata tetraploids.

Under salt stress, recretohalophyte Plumbago auriculata tetraploids enhance salt tolerance by increasing selective secretion of Na+ compared with that in diploids, although the mechanism is unclear. Using non-invasive micro-test technology, the effect of salt gland Ca2+ content on Na+ and K+ secretion were investigated in diploid and tetraploid P. auriculata under salt stress. Salt gland Ca2+ content and secretion rates of Na+ and K+ were higher in tetraploids than in diploids under salt stress. Addition of exogenous Ca2+ increased the Ca2+ content of the salt gland in diploids and is accompanied by an increase in the rate of Na+ and K+ secretion. With addition of a Ca2+ channel inhibitor, diploid salt glands retained large amounts of Ca2+, leading to higher Ca2+ content and Na+ secretion rate than those of tetraploids. Inhibiting H2O2 generation and H+-ATPase activity altered Na+ and K+ secretion rates in diploids and tetraploids under salt stress, indicating involvement in regulating Na+ and K+ secretion. Our results indicate that the increased Na+ secretion rate of salt gland in tetraploids under salt stress was associated with elevated Ca2+ content in salt gland.

Comparative stigmatic transcriptomics reveals self and cross pollination responses to heteromorphic incompatibility in Plumbago auriculata Lam.

"Heteromorphic self-incompatibility" (HetSI) in plants is a mechanism of defense to avoid self-pollination and promote outcrossing. However, the molecular mechanism underlying HetSI remains largely unknown. In this study, RNA-seq was conducted to explore the molecular mechanisms underlying self-compatible (SC, "T × P" and "P × T") and self-incompatible (SI, "T × T" and "P × P") pollination in the two types of flowers of Plumbago auriculata Lam. which is a representative HetSI plant. By comparing "T × P" vs. "T × T", 3773 (1407 upregulated and 2366 downregulated) differentially expressed genes (DEGs) were identified, 1261 DEGs between "P × T" and "P × P" (502 upregulated and 759 downregulated). The processes in which these DEGs were significantly enriched were "MAPK (Mitogen-Activated Protein Kinases-plant) signaling pathway", "plant-pathogen interaction","plant hormone signal transduction", and "pentose and glucuronate interconversion" pathways. Surprisingly, we discovered that under various pollination conditions, multiple notable genes that may be involved in HetSI exhibited distinct regulation. We can infer that the HetSI strategy might be unique in P. auriculata. It was similar to "sporophytic self-incompatibility" (SSI) but the HetSI mechanisms in pin and thrum flowers are diverse. In this study, new hypotheses and inferences were proposed, which can provide a reference for crop production and breeding.

Methyl jasmonate regulation of pectin polysaccharides in Cosmos bipinnatus roots: A mechanistic insight into alleviating cadmium toxicity.

Methyl jasmonate (MeJA), a crucial phytohormone, which plays an important role in resistance to Cadmium (Cd) stress. The cell wall (CW) of root system is the main location of Cd and plays a key role in resistance to Cd toxicity. However, the mechanism effect of MeJA on the CW composition and Cd accumulation remain unclear. In this study, the contribution of MeJA in regulating CW structure, pectin composition and Cd accumulation was investigated in Cosmos bipinnatus. Phenotypic results affirm MeJA's significant role in reducing Cd-induced toxicity in C. bipinnatus. Notably, MeJA exerts a dual impact, reducing Cd uptake in roots while increasing Cd accumulation in the CW, particularly bound to pectin. The molecular structure of pectin, mainly uronic acid (UA), correlates positively with Cd content, consistent in HC1 and cellulose, emphasizing UA as pivotal for Cd binding. Furthermore, MeJA modulates pectin methylesterase (PME) activity under Cd stress, influencing pectin's molecular structure and homogalacturonan (HG) content affecting Cd-binding capacity. Chelate-soluble pectin (CSP) within soluble pectins accumulates a substantial Cd proportion, with MeJA regulating both UA content and the minor component 3-deoxy-oct-2-ulosonic acid (Kdo) in CSP. The study delves into the intricate regulation of pectin monosaccharide composition under Cd stress, revealing insights into the CW's physical defense and Cd binding. In summary, this research provides novel insights into MeJA-specific mechanisms alleviating Cd toxicity in C. bipinnatus, shedding light on complex interactions between MeJA, and Cd accumulation in CW pectin polysaccharide.

ISSR molecular markers and anatomical structures can assist in rapid and directional screening of cold-tolerant seedling mutants of medicinal and ornamental plant in Plumbago indica L.

Plumbago indica L. is a perennial herb with ornamental and anticancer medicinal functions widely distributed in the tropics. It is affected by temperature and cannot bloom normally in colder subtropical regions, which seriously affects its ornamental value. To create low-temperature resistance mutants and enrich new germplasm resources, this study used tissue culture and chemical reagent (0.5 mmol/L NaN3) and low-temperature stress (0°C, full darkness for 48h) induction to target and screen for cold-resistance mutants. The results showed that the ISSR band polymorphism ratio of the 24 suspected mutant materials was 87.5%. The DNA profiles of the 9 mutants initially identified were altered. The content of plumbagin in the stems and leaves of the mutants was examined, and it was found that the accumulation in the leaves of the mutant SA24 could be as high as 3.84 times that of the control, which was 0.5991%. There were significant differences in the anatomical structures of roots, stems and leaves. The mutants mostly exhibited reduced root diameter (only 0.17-0.69 times that of CK), increased stem diameter (up to 2.19 times that of CK), enlarged mesophyll cells, increased thickness (up to 1.83 times that of CK) and high specificity, which are thought to be important for the different cold resistance obtained by the mutants. In the cold resistance experiment, four cold-tolerant mutants were successfully screened according to their morphological characteristics and physiological indexes, and the mutagenesis efficiency could be as high as 2.22% and did not affect the accumulation of plumbagin in their stems and leaves, even higher than CK. The responses of the screened mutants SA15, SA19, SA23 and SA24 to low temperature showed slower leaf wilting, higher light energy conversion efficiency, less accumulation of MDA content, increased enzymatic activities of antioxidant enzymes (SOD, CAT, POD) and more accumulation of soluble sugars and proline content. These characteristics are consistent with the response of cold-resistance plants to low temperatures. The cold- resistance mutants cultivated in soil were observed of agronomic and ornamental traits for one year, mainly manifested as delayed flowering and delayed entry into the senescence stage. This study provides a more rapid and accurate technique for identifying and screening cold-tolerant mutants, and lays the foundation for future experiments on the creation of new cold-resistant varieties.

Abscisic acid (ABA) alleviates cadmium toxicity by enhancing the adsorption of cadmium to root cell walls and inducing antioxidant defense system of Cosmos bipinnatus.

Cadmium (Cd) pollution is a global problem affecting soil ecology and plant growth. Abscisic acid (ABA) acts as a growth and stress hormone, regulates cell wall synthesis, and plays an important role in plant responses to stress. There are few studies on the mechanisms behind abscisic acid alleviation of cadmium stress in Cosmos bipinnatus, especially in regards to regulation of the root cell wall. This study examined the effects of different concentrations of abscisic acid at different concentrations of cadmium stress. Through adding 5 µmol/L and 30 µmol/L cadmium, followed by spraying 10 µmol/L and 40 µmol/L ABA in a hydroponic experiment, it was found that under two concentrations of cadmium stress, low concentration of ABA improved root cell wall polysaccharide, Cd, and uronic acid content. Especially in pectin, after the application of low concentration ABA, the cadmium concentration was significantly increased by 1.5 times and 1.2 times compared with the Cd concentration under Cd5 and Cd30 treatment alone, respectively. Fourier-Transform Infrared spectroscopy (FTIR) demonstrated that cell wall functional groups such as -OH and -COOH were increased with exposure to ABA. Additionally, the exogenous ABA also increased expression of three kinds of antioxidant enzymes and plant antioxidants. The results of this study suggest that ABA could reduce Cd stress by increasing Cd accumulation, promoting Cd adsorption on the root cell wall, and activating protective mechanisms. This result could help promote application of C. bipinnatus for phytostabilization of cadmium-contaminated soil.

Inheritance of distyly and homostyly in self-incompatible Primula forbesii.

The evolutionary transition from self-incompatible distyly to self-compatible homostyly frequently occurs in heterostylous taxa. Although the inheritance of distyly and homostyly has been deeply studied, our understanding on modifications of the classical simple Mendelian model is still lacking. Primula forbesii, a biennial herb native to southwest China, is a typical distylous species, but after about 20 years of cultivation with open pollination, self-compatible homostyly appeared, providing ideal material for the study of the inheritance of distyly and homostyly. In this study, exogenous homobrassinolide was used to break the heteromorphic incompatibility of P. forbesii. Furthermore, we performed artificial pollination and open-pollination experiments to observe the distribution of floral morphs in progeny produced by different crosses. The viability of seeds from self-pollination was always the lowest among all crosses, and the homozygous S-morph plants (S/S) occurred in artificial pollination experiments but may experience viability selection. The distyly of P. forbesii is governed by a single S-locus, with S-morph dominant hemizygotes (S/-) and L-morph recessive homozygotes (-/-). Homostylous plants have a genotype similar to L-morph plants, and homostyly may be caused by one or more unlinked modifier genes outside the S-locus. Open pollinations confirm that autonomous self-pollination occurs frequently in L-morphs and homostylous plants. This study deepens the understanding of the inheritance of distyly and details a case of homostyly that likely originated from one or more modifier genes.

Enhanced Na+ and Cl- sequestration and secretion selectivity contribute to high salt tolerance in the tetraploid recretohalophyte Plumbago auriculata Lam.

MAIN CONCLUSION: Enhanced secretion of Na+ and Cl- in leaf glands and leaf vacuolar sequestration of Na+ or root retention of Cl-, combined with K+ retention, contribute to the improved salt tolerance of tetraploid recretohalophyte P. auriculata. Salt stress is one of the major abiotic factors threatening plant growth and development, and polyploids generally exhibit higher salt stress resistance than diploids. In recretohalophytes, which secrete ions from the salt gland in leaf epidermal cells, the effects of polyploidization on ion homeostasis and secretion remain unknown. In this study, we compared the morphology, physiology, and ion homeostasis regulation of diploid and autotetraploid accessions of the recretohalophyte Plumbago auriculata Lam. after treatment with 300 mM NaCl for 0, 2, 4, 6, and 8 days. The results showed that salt stress altered the morphology, photosynthetic efficiency, and chloroplast structure of diploid P. auriculata to a greater extent than those of its tetraploid counterpart. Moreover, the contents of organic osmoregulatory substances (proline and soluble sugars) were significantly higher in the tetraploid than in the diploid, while those of H2O2 and malondialdehyde (MDA) were significantly lower. Analysis of ion homeostasis revealed that the tetraploid cytotype accumulated more Na+ in stems and leaves and more Cl- in roots but less K+ loss in roots compared with diploid P. auriculata. Additionally, the rate of Na+ and Cl- secretion from the leaf surface was higher, while that of K+, Mg2+, and Ca2+ secretion was lower in tetraploid plants. X-ray microanalysis of mesophyll cells revealed that Na+ mainly accumulated in different cellular compartments in the tetraploid (vacuole) and diploid (cytoplasm) plants. Our results suggest that polyploid recretohalophytes require the ability to sequester Na+ and Cl-(via accumulation in leaf cell vacuoles or unloading by roots) and selectively secrete these ions (through salt glands) together with the ability to prevent K+ loss (by roots). This mechanism required to maintain K+/Na+ homeostasis in polyploid recretohalophytes under high salinity provides new insights in the improved maintenance of ion homeostasis in polyploids under salt stress.

Effect of chromium stress on metal accumulation and cell wall fractions in Cosmos bipinnatus.

As one of the major pollutants in the environment, chromium (Cr), a heavy metal, poses a serious threat to urban green spaces and human life and health. Cosmos bipinnatus is considered a potential accumulator of Cr, and the differences in cellular Cr distribution and compartmentalization may uncover the mechanisms involved in its tolerance to Cr. To elucidate the effects of Cr stress on C. bipinnatus and determine the mechanism of Cr tolerance in C. bipinnatus, we investigated the physiological indicators, subcellular distribution and chemical forms, cell wall fractions and their Cr contents, uronic acid content in the cell wall fractions, and Fourier transform infrared spectroscopy (FTIR) of the cell wall. The results showed that the antioxidant enzyme activities in C. bipinnatus under Cr stress and most of the Cr were fixed in the cell wall. Notably, changes in the content of pectin fractions in the cell wall affected the accumulation of Cr in the cell wall of C. bipinnatus and the stability of negatively charged groups. In addition, the carboxyl and hydroxyl groups played a role in fixing metal in various parts of the C. bipinnatus cell wall.

Gas chromatography-mass spectrometry analysis of floral fragrance-related compounds in scented rose (Rosa hybrida) varieties and a subsequent evaluation on the basis of the analytical hierarchy process.

Scented rose (Rosa hybrida) varieties are valued as ornamentals, but they also contain volatile organic compounds (VOCs) that produce pleasant aromas. In plants, aromas are produced via metabolism during growth, and each aroma compound has a unique function. In this study, the floral aroma compounds of diverse scented rose varieties were analyzed and classified. The VOCs of different rose varieties were qualitatively and quantitatively analyzed via headspace solid-phase microextraction combined with gas chromatography and mass spectrometry. The test materials were the mature flowers of 55 scented rose varieties that were cultivated under identical conditions. Seventeen important aroma compounds were selected and an analytical hierarchy process (AHP)-based method was developed to identify the most suitable essential oil resources, aromatherapy resources, and healthcare resources. A floral fragrance evaluation model was established for the comprehensive evaluation of the scented rose varieties. The 55 varieties were classified into three grades according to their suitability for each use. 'Soeur Emmanuelle', 'Wollerton Old Hall', 'Accademia', and 'Tianmidemeng' were revealed to be suitable essential oil, aromatherapy, and healthcare resources. On the basis of their aroma compound types, the fifty-five rose varieties were divided into eight groups. The results of this study provide the theoretical basis for the classification of rose flower aromas as well as the rational use of diverse rose varieties to further develop the rose industry.

Efficiency of heterogeneous chelating agents on the phytoremediation potential and growth of Sasa argenteostriata (Regel) E.G. Camus on Pb-contaminated soil.

Ethylenediaminetetraacetic acid (EDTA) is one of the most effective chelating agents for enhancing lead (Pb) accumulation in various plant organs. However, it has a higher risk of causing secondary pollution than other chelating agents. To reduce such environmental risks and increase remediation efficiency, EDTA can be combined with degradable chelating agents for use in phytoremediation, but there are few reports on the combination of EDTA and nitrilotriacetic acid (NTA). This study evaluated the effects of combined EDTA and NTA application at different concentrations (900, 1200, or 1500 mg/kg) and with different methods (1 application or 3 applications) on dwarf bamboo (Sasa argenteostriata (Regel) E.G. Camus) growth and phytoremediation efficiency and on the soil environment in pot experiments with Pb-contaminated soil. Applying EDTA and NTA together resulted in lower soil water-soluble Pb concentrations than applying EDTA alone and therefore resulted in lower environmental risk. The increased availability of soil Pb produced a stress response in the dwarf bamboo plants, which increased their biomass significantly. Moreover, under the chelating treatments, the soil Pb availability increased, which promoted Pb translocation in plants. The Pb content in the aerial parts of the dwarf bamboo increased significantly in all treatments (translocation factors increased by 300~1500% compared with that in CK). The Pb content increase in the aerial parts caused high proline accumulation in dwarf bamboo leaves, to alleviate Pb toxicity. Maximum Pb accumulation was observed in the EN1500 treatment, which was significantly higher than that in the other treatments except the EN900 treatment. This study elucidates the choice of remediation techniques and the physiological characteristics of the plants used in such studies. In conclusion, the EN900 treatment resulted in the lowest environmental risk, greatest biomass production, and highest phytoremediation efficiency of all treatments, indicating that it has great potential for application in phytoremediation with dwarf bamboo in Pb-contaminated soil.

Transcriptome Analysis Revealed a Cold Stress-Responsive Transcription Factor, PaDREB1A, in Plumbago auriculata That Can Confer Cold Tolerance in Transgenic Arabidopsis thaliana.

The tropical plant Plumbago auriculata can tolerate subzero temperatures without induction of apoptosis after cold acclimation in autumn, making it more cold tolerant than conventional tropical plants. In this study, we found that low temperatures significantly affected the photosynthetic system of P. auriculata. Using transcriptome sequencing, PaDREB1A was identified as a key transcription factor involved in the response to cold stress in P. auriculata. This transcription factor may be regulated by upstream JA signaling and regulates downstream ERD4 and ERD7 expression to resist cold stress. Overexpression of PaDREB1A significantly enhanced freezing resistance, protected the photosynthetic system, and enhanced the ROS scavenging mechanism under cold stress in Arabidopsis thaliana. Additionally, PaDREB1A significantly enhanced the expression of CORs and CAT1 in A. thaliana, which further activated the downstream pathway to enhance plant cold tolerance. This study explored the possible different regulatory modes of CBFs in tropical plants and can serve as an important reference for the introduction of tropical plants to low-temperature regions.

Dynamic study of the lead (Pb) tolerance and accumulation characteristics of new dwarf bamboo in Pb-contaminated soil.

Dwarf bamboo is a woody plant with potential for use in the remediation of Pb-contaminated soil. Due to its clonal growth habit, there are two keys to its application for continuous soil Pb remediation: 1) its ability to form shoots and grow into new bamboo normally under Pb stress and 2) the Pb tolerance and accumulation characteristics of this new bamboo. Here, 5 species of dwarf bamboo were treated with 2 levels of soil Pb stress (0 and 1500 mg kg-1). In the roots of 3 of the species (Sasa argenteostriata, Sasaella glabra, and Indocalamus decorus), Pb tended to be distributed along the cell wall and transported to vacuoles. In the other 2 species (Sasa auricoma and Sasa fortunei), Pb was arranged linearly along the cell wall. Under Pb treatment, the new bamboo of all species showed gradual physiological adaptation to Pb stress. Correlations of the net photosynthetic rate, superoxide dismutase activity, and free proline levels with Pb content in new leaves in November were all higher than those in July, though that of malondialdehyde content decreased, suggesting that new dwarf bamboo exhibits good soil Pb stress tolerance. Sasa argenteostriata and Indocalamus decorus consistently maintained higher antioxidant enzyme activities and free proline levels than the other species under Pb treatment, and the total biomass per pot of the new bamboo decreased the least compared to that in the Pb-free treatment for these two species. Therefore, these bamboo species may be used in the long-term continuous remediation of Pb-contaminated soil.

The nonreciprocal heterostyly and heterotypic self-incompatibility of Ceratostigma willmottianum.

Ceratostigma willmottianum (Plumbaginaceae) is a perennial herb native to China. Many species of Plumbaginaceae have been reported to exhibit heterostyly. Determining the functional breeding system of C. willmottianum can improve our understanding of the reproductive ecology of heterostylous plants. We investigated the floral traits and pollen and stigma characteristics in a natural population, and artificial pollination was carried out in an artificial population. It was found that C. willmottianum was distylous with short (S)- and long (L)-styled morphs, did not exhibit precise reciprocal herkogamy and was partially self-compatible but primarily outcrossing. In the artificial pollination experiments, the pollen tubes reached the base of the style under intermorph pollination, whereas they rarely penetrated the style under intramorph pollination and self-pollination. Both the L and S morphs exhibited a high seed set after intermorph pollination, whereas the seed set under intramorph pollination was lower. Therefore, C. willmottianum may spread the chance of receiving pollen between the two morphs by nonreciprocal heterostyly, which may be a unique mode of ecological adaptation in Plumbaginaceae. We believe that our discovery could provide new ideas regarding the origin and evolution of heterostyly.

The presence of zinc reduced cadmium uptake and translocation in Cosmos bipinnatus seedlings under cadmium/zinc combined stress.

Cadmium (Cd) and zinc (Zn) coexist in the environment but interact differently in plants. Cosmos bipinnatus has been potentially considered as a Cd-accumulator. Thus, this study investigated the detoxification mechanism in C. bipinnatus seedlings under Cd, Zn and Cd + Zn stresses. In the present study, the presence of Zn inhibited Cd uptake and translocation, whereas Cd merely hindered Zn uptake. The concentration of Cd in soluble fraction significantly decreased and Cd was bounded to the cell wall in root under Cd + Zn stress. Meanwhile, Zn and Cd mutually decreased their concentrations in the ethanol extractable form (FE) and water extractable form (FW) in roots and shoots. Furthermore, Cd + Zn stress enhanced the activities of superoxide dismutase (SOD, EC 1.15.1.1), peroxidase (POD, EC 1.11.1.7) and catalase (CAT, EC 1.11.1.6) compared to Cd stress alone. These results suggested that Zn effectively decreased Cd uptake and translocation, changed their subcellular distributions, regulated their chemical forms composition and increased antioxidative enzyme activities, thereby enhancing the tolerance to Cd in C. bipinnatus. This study physiologically revealed the interactive effect of Cd and Zn on the detoxification mechanism of Cd in C. bipinnatus and provided new information on phytoremediation of the heavy metal contaminated soils.

Different strategies for lead detoxification in dwarf bamboo tissues.

Dwarf bamboo Sasa argenteostriata (Regel) E.G. Camus is considered as potential plants for metal phytoremediation in previous filed observations. However, the mechanisms of lead (Pb) detoxification has not been described. The objective of this study was to explore the difference strategies or mechanisms of Pb detoxification in plant tissues. In this regard, four Pb treatments with hydroponics including 0 (control), 300, 600, and 900 mg L-1 were conducted to examine subcellular compartmentalization, Pb accumulation/species and antioxidant-assisted chelation. Our findings showed the retention of Pb by the whip-root system is one of its detoxification mechanisms to avoid damage the shoots. In addition, the cell wall retention is the dominant detoxification strategy of whips, new roots, old roots and new/old stems, while vacuolar compartmentalization is for new/old leaves. Interestingly, four low-mobility/-toxicity Pb species (i.e., FNaCl, FHAc, FHCl and FR) are distributed in roots, whips and stems, while two high-mobility/-toxicity Pb species (FE and FW) in leaves. The conversion of Pb to low-toxicity/-migration is a Pb-detoxification strategy in roots, whips and stems but not in leaves. Besides, the new/old roots and leaves can alleviate Pb damage through the synthesis of non-protein thiol, glutathione and phytochelatins. Among these, phytochelatins play a leading role in the detoxification in new/old roots, while glutathione is in new/old leaves. This study provides the first comprehensive evidence regarding the different strategies for Pb detoxification in dwarf bamboo tissues from physiological to cellular level, supporting that this plant could be rehabilitated for phytoremediation in Pb-contaminated media.

Untargeted metabolomic profiling reveals that different responses to self and cross pollination in each flower morph of the heteromorphic plant Plumbago auriculata.

Heteromorphic self-incompatibility (HetSI), which is regulated by sporophytes, occurs in some species as a strategy to promote cross-pollination. This research aimed to reveal metabolic changes occurring in HetSI. We used fluorescence microscopy as a tool to compare growth behavior in self-incompatible (SI) and self-compatible (SC) pollination in both pin and thrum flowers of Plumbago auriculata and to identify the ideal timepoint for sample collection for subsequent experiments. We also employed scanning electron microscopy (SEM) to evaluate intermorph structural differences in the pollen grains and stigmas in relation to HetSI. Importantly, UPLC-MS/MS was applied in this study to identify metabolites, compare metabolic differences between pin and thrum styles and monitor metabolic changes in SC and SI pollinations in the two types of flowers. The metabolites mainly included amino acids/peptides, flavonoids, glycosides/sugars, phenols, other organic acids, fatty acids (derivatives)/lipids, amines, aldehydes, alkaloids, alcohols and other compounds. Surprisingly, energy-related nutrients such as amino acids/peptides and tricarboxylic acid cycle-related metabolites were found at higher levels in SI pollinations than in SC pollinations. This result indicates that physiological changes in pollen-stigma interactions differ in pin and thrum styles and SC and SI pollinations and that energy deficiency is not one of the reasons for HetSI.

Physiological and transcriptome response to cadmium in cosmos (Cosmos bipinnatus Cav.) seedlings.

To date, several species of Asteraceae have been considered as Cd-accumulators. However, little information on the Cd tolerance and associated mechanisms of Asteraceae species Cosmos bipinnatus, is known. Presently, several physiological indexes and transcriptome profiling under Cd stress were investigated. C. bipinnatus exhibited strong Cd tolerance and recommended as a Cd-accumulator, although the biomasses were reduced by Cd. Meanwhile, Cd stresses reduced Zn and Ca uptake, but increased Fe uptake. Subcellular distribution indicated that the vacuole sequestration in root mainly detoxified Cd under lower Cd stress. Whilst, cell wall binding and vacuole sequestration in root co-detoxified Cd under high Cd exposure. Meanwhile, 66,407 unigenes were assembled and 41,674 (62.75%) unigenes were annotated in at least one database. 2,658 DEGs including 1,292 up-regulated unigenes and 1,366 down-regulated unigenes were identified under 40 µmol/L Cd stress. Among of these DEGs, ZIPs, HMAs, NRAMPs and ABC transporters might participate in Cd uptake, translocation and accumulation. Many DEGs participating in several processes such as cell wall biosynthesis, GSH metabolism, TCA cycle and antioxidant system probably play critical roles in cell wall binding, vacuole sequestration and detoxification. These results provided a novel insight into the physiological and transcriptome response to Cd in C. bipinnatus seedlings.

The evolution and utility of ribosomal ITS sequences in Bambusinae and related species: divergence, pseudogenes, and implications for phylogeny.

Ribosomal internal transcribed spacer (ITS) sequences are commonly used for phylogenetic reconstruction because they are highly reiterated as components of rDNA repeats, and hence are often subject to rapid homogenization through concerted evolution. Concerted evolution leads to intragenomic uniformity of repeats even between loci on nonhomologous chromosomes. However, a number of studies have shown that the ITS polymorphism within individuals is quite common. The molecular systematics of Bambusinae and related species were recently assessed by different teams using independently generated ITS sequences, and the results disagreed in some remarkable features. Here we compared the ITS sequences of the members of Bambusa s. l., the genera Dendrocalamus, Dinochloa, Gigantochloa, Guadua, Melocalamus, Monocladus, Oxytenanthera, Thyrsostachys, Pleioblastus, Pseudosasa and Schizostachyum.We have reanalysed the ITS sequences used by different research teams to reveal the underlying patterns of their different results. After excluding the sequences suspected to represent paralogous loci, a phylogenetic analysis of the subtribe Bambusinae species were performed using maximum parsimony and maximum-likelihood methods. The implications of the findings are discussed. The risk of incorporating ITS paralogues in plant evolutionary studies that can distort the phylogenetic signal should caution molecular systematists.