Association among starch storage, metabolism, related genes and growth of Moso bamboo (Phyllostachys heterocycla) shoots

Background Both underground rhizomes/buds and above-ground Moso bamboo (Phyllostachys heterocycla) shoots/culms/branches are connected together into a close inter-connecting system in which nutrients are transported and shared among each organ. However, the starch storage and utilization mechanisms during bamboo shoot growth remain unclear. This study aimed to reveal in which organs starch was stored, how carbohydrates were transformed among each organ, and how the expression of key genes was regulated during bamboo shoot growth and developmental stages which should lay a foundation for developing new theoretical techniques for bamboo cultivation. Results Based on changes of the NSC content, starch metabolism-related enzyme activity and gene expression from S0 to S3, we observed that starch grains were mainly elliptical in shape and proliferated through budding and constriction. Content of both soluble sugar and starch in bamboo shoot peaked at S0, in which the former decreased gradually, and the latter initially decreased and then increased as shoots grew. Starch synthesis-related enzymes (AGPase, GBSS and SBE) and starch hydrolase (α-amylase and β-amylase) activities exhibited the same dynamic change patterns as those of the starch content. From S0 to S3, the activity of starch synthesis-related enzyme and starch amylase in bamboo rhizome was significantly higher than that in bamboo shoot, while the NSC content in rhizomes was obviously lower than that in bamboo shoots. It was revealed by the comparative transcriptome analysis that the expression of starch synthesis-related enzyme-encoding genes were increased at S0, but reduced thereafter, with almost the same dynamic change tendency as the starch content and metabolism-related enzymes, especially during S0 and S1. It was revealed by the gene interaction analysis that AGPase and SBE were core genes for the starch and sucrose metabolism pathway. Conclusions Bamboo shoots were the main organ in which starch was stored, while bamboo rhizome should be mainly functioned as a carbohydrate transportation channel and the second carbohydrate sink. Starch metabolism-related genes were expressed at the transcriptional level during underground growth, but at the post-transcriptional level during above-ground growth. It may be possible to enhance edible bamboo shoot quality for an alternative starch source through genetic engineering.

Microwave-assisted extraction of flavonoids from Phyllostachys heterocycla leaves: Optimization, mechanism, and antioxidant activity in vitro

Flavonoids were extracted from Phyllostachys heterocycla leaves by adopting microwave-assisted extraction technology. Based on the single factor experiment and Plackett-Burman design results, the extraction process of flavonoids was further optimized using the response surface methodology. The optimum conditions were as follows: an ethanol concentration of 78.1%, an extraction time of 24.9 min, and a microwave power of 559 W. Under these conditions, the extraction yield of flavonoids was 4.67%, which was in close proximity to the predicted value (4.70%) and higher than the extraction yield from traditional Soxhlet extraction (3.35%). Moreover, the possible extraction mechanisms of these two extraction methods were further derived to explain why the microwave-assisted extraction of flavonoids was more efficient compared with traditional Soxhlet extraction. Ultimately, the antioxidant activities in vitro of flavonoids from Phyllostachys heterocycla leaves were evaluated via DPPH and ABTS radical scavenging assay. The flavonoids from Phyllostachys heterocycla leaves exhibited excellent antioxidant activities in vitro and Phyllostachys heterocycla leaves could be a new natural source for developing antioxidants. Overall, the findings of this research could provide a theoretical reference for the further comprehensive development and utilization of bamboo resources.

Chemical Constituent Profiling of Phyllostachys heterocycla var. Pubescens with Selective Cytotoxic Polar Fraction through EGFR Inhibition in HepG2 Cells

Different extracts of the Bamboo shoot skin Phyllostachys heterocycla var. pubescens were screened against panel of cancer cell lines and normal one. The cell viability results exhibited that the ethyl acetate extract showed the least vitality percentage of 2.14% of HepG2 cells. Accordingly, it was subjected to chromatographic separation, which resulted in the isolation of a new natural product; 7-hydroxy, 5-methoxy, methyl cinnamate (1), together with four known compounds. The structures of the pure isolated compounds were deduced based on different spectroscopic data. The new compound (1) was screened against the HepG2 and MCF-7 cells and showed IC50 values of 7.43 and 10.65 µM, respectively. It induced apoptotic cell death in HepG2 with total apoptotic cell death of 58.6% (12.44-fold) compared to 4.71% in control by arresting cell cycle progression at the G1 phase. Finally, compound 1 was validated as EGFR tyrosine kinase inhibitor in both enzymatic levels (IC50 = 98.65 nM compared to Erlotinib (IC50 = 78.65 nM). Finally, in silico studies of compound 1 through the molecular docking indicated its high binding affinity towards EGFR protein and the ADME pharmacokinetics indicated it as a drug-like.

Effects of Different Management Practices on the Increase in Phytolith-Occluded Carbon in Moso Bamboo Forests

Phytolith-occluded carbon (PhytOC), a promising long-term biogeochemical carbon sequestration mode, plays a crucial role in the global carbon cycle and the regulation of atmospheric CO2. Previous studies mostly focused on the estimation of the content and storage of PhytOC, while it remains unclear about how the management practices affect the PhytOC content and whether it varies with stand age. Moso bamboo (Phyllostachys heterocycla var. pubescens) has a great potential in carbon sequestration and is rich in PhytOC. Here, we selected four management treatments, including control (CK), compound fertilization (CF), silicon (Si) fertilization (SiF) (monosilicic acid can form phytoliths through silicification), and cut to investigate the variation of phytoliths and PhytOC contents in soil, leaves, and litters, and their storage in Moso bamboo forests. In soil, the SiF fertilizer treatment significantly (P < 0.05) increased phytolith content, PhytOC content, and storage compared to CK, while there were no significant differences between the treatments of CF and cut. In leaf, compared with CK, phytolith content of the second-degree leaves under SiF and the first-degree leaves under cut treatment significantly increased, and the three treatments significantly increased PhytOC storage for leaves with three age classes. In litter, the phytolith and PhytOC contents under the three treatments were not significantly different from that under the CK treatment. The PhytOC storage increased by 19.33% under SiF treatment, but significantly decreased by 40.63% under the CF treatment. For the entire Moso bamboo forest ecosystems, PhytOC storage of all the three management treatments increased compared with CK, with the largest increase by 102% under the SiF treatment. The effects of management practices on the accumulation of PhytOC varied with age. Our study implied that Si fertilization has a greater potential to significantly promote the capacity of sequestration of carbon in Moso bamboo forests.

Cytotoxic, Apoptosis-Inducing Activities, and Molecular Docking of a New Sterol from Bamboo Shoot Skin Phyllostachys heterocycla var. pubescens

Phytochemical screening of nonpolar fractions from the methanol extract of the Bamboo shoot skin Phyllostachys heterocycla var. pubescens resulted in the isolation of a new sterol-glucoside-fatty acid derivative (6’-O-octadeca-8″,11″-dienoyl)-sitosterol-3-O-β-d-glucoside (1), together with six known compounds. The chemical structures of the pure isolated compounds were deduced based on different spectral data. The isolated compounds were assessed to determine their cytotoxic activity, and the results were confirmed by determining their apoptotic activity. Compound 1 was more cytotoxic against the MCF-7 cells (IC50 = 25.8 µM) compared to Fluorouracil (5-FU) (26.98 µM), and it significantly stimulated apoptotic breast cancer cell death with 32.6-fold (16.63% compared to 0.51 for the control) at pre-G1 and G2/M-phase cell cycle arrest and blocked the progression of MCF-7 cells. Additionally, RT-PCR results further confirmed the apoptotic activity of compound 1 by the upregulation of proapoptotic genes (P53; Bax; and caspases 3, 8, and 9) and downregulation of the antiapoptotic genes (BCL2). Finally, the identified compounds, especially 1, were found to have high binding affinity towards both tyrosine-specific protein kinase (TPK) and vascular endothelial growth factor receptor (VEGFR-2) through the molecular docking studies that highlight its mode of action.

Superior growth, N uptake and NH4+ tolerance in the giant bamboo Phyllostachys edulis over the broad-leaved tree Castanopsis fargesii at elevated NH4+ may underlie community succession and favor the expansion of bamboo

The unbridled expansion of bamboo has imposed serious threats on ecosystem processes and functions. Considerable evidence indicates that bamboo invasions can alter plant-available soil nitrogen (N) pools and rates of N cycling, but the consequences of altered N availability for plant growth and community structure have thus far been poorly characterized. The primary soil-accessible N forms for most plants are ammonium (NH4+) and nitrate (NO3−), but plants differ in their ability to use the different N forms, and these differences can be related to their ecological characteristics and drive community structure. In this context, we evaluated the growth response, N uptake and interspecific competition in two subtropical species, Phyllostachys edulis (Carrière) J. Houzeau (Synonym Phyllostachys heterocycla Carrière) and Castanopsis fargesii Franch., dominant species of bamboo and secondary evergreen broad-leaved forests, respectively, under changing N availability in seedlings supplied with different N concentrations and NH4+/NO3− proportions, in vermiculite culture, in a controlled environment. The results show that (i) both species display an NH4+ preference at elevated N concentrations. The growth of P. edulis seedlings was strongly enhanced at increased ratios of NH4+ to NO3− especially at higher N concentrations, but to a much lesser extent in C. fargesii. (ii) NH4+ preference at the level of N uptake in both species was confirmed by the Non-invasive Micro-test Technology and by examining 15N signatures. Phyllostachys edulis had higher NH4+ net fluxes and N concentration under NH4+ treatments than C. fargesii. (iii) NH4+ at higher concentrations caused toxicity in both species as it inhibited root growth and even caused seedling death, but P. edulis had a higher NH4+-tolerance threshold (24 mM) than C. fargesii (16 mM). (iv) When mixed-species cultures were examined in an NH4+-rich medium, the growth of C. fargesii, but not P. edulis, was significantly inhibited compared with growth in monoculture. Therefore, P. edulis exhibited stronger plasticity and adaptation to changing N availability, whereas C. fargesii had low responsiveness and capacity to acclimate to soil N changes. Phyllostachys edulis displayed a significant competitive growth advantage compared with C. fargesii on NH4+-dominated substrates.

The acid invertase gene family is involved in internode elongation in Phyllostachys heterocycla cv. pubescens

Acid invertases (INVs) play a pivotal role in both vegetative and reproductive growth of plants. However, their possible functions in fast-growing plants such as bamboo are largely unknown. Here, we report the molecular characterization of acid INVs in Phyllostachys heterocycla cv. pubescens, a fast-growing bamboo species commercially grown worldwide. Nine acid INVs (PhINVs), including seven cell wall INVs (PhCWINV1, PhCWINV2, PhCWINV3, PhCWINV4, PhCWINV5, PhCWINV6 and PhCWINV7) and two vacuolar INVs (PhVINV11 and PhVINV12) were isolated. Bioinformatic analyses demonstrated that they all share high amino acid identity with other INVs from different plant species and contain the motifs typically conserved in acid INV. Enzyme activity assays revealed a significantly higher INV activity in the fast-growing tissues, such as the elongating internodes of stems. Detailed quantitative reverse-transcription PCR analyses showed various expression patterns of PhINVs at different developmental stages of the elongating stems. With the exception of PhCWINV6, all PhINVs were ubiquitously expressed in a developmental-specific manner. Further studies in Arabidopsis exhibited that constitutive expression of PhCWINV1, PhCWINV4 or PhCWINV7 increased the biomass production of transgenic plants, as indicated by augmented plant heights and shoot dry weights than the wild-type plants. All these results suggest that acid INVs play a crucial role in the internode elongation of P. heterocycla cv. pubescens and would provide valuable information for the dissection of their exact biological functions in the fast growth of bamboo.