Integrated Analysis of Transcriptome and Metabolome Reveals the Regulation of Chitooligosaccharide on Drought Tolerance in Sugarcane (Saccharum spp. Hybrid) under Drought Stress.

Sugarcane (Saccharum spp. hybrid) is an important crop for sugar and biofuels, and often suffers from water shortages during growth. Currently, there is limited knowledge concerning the molecular mechanism involved in sugarcane response to drought stress (DS) and whether chitooligosaccharide could alleviate DS. Here, we carried out a combined transcriptome and metabolome of sugarcane in three different treatment groups: control group (CG), DS group, and DS + chitooligosaccharide group (COS). A total of 12,275 (6404 up-regulated and 5871 down-regulated) differentially expressed genes (DEGs) were identified when comparing the CG and DS transcriptomes (T_CG/DS), and 2525 (1261 up-regulated and 1264 down-regulated) DEGs were identified in comparing the DS and COS transcriptomes (T_DS/COS). GO and KEGG analysis showed that DEGs associated with photosynthesis were significantly enriched and had down-regulated expression. For T_DS/COS, photosynthesis DEGs were also significantly enriched but had up-regulated expression. Together, these results indicate that DS of sugarcane has a significantly negative influence on photosynthesis, and that COS can alleviate these negative effects. In metabolome analysis, lipids, others, amino acids and derivatives and alkaloids were the main significantly different metabolites (SDMs) observed in sugarcane response to DS, and COS treatment reduced the content of these metabolites. KEGG analysis of the metabolome showed that 2-oxocarboxylic acid metabolism, ABC transporters, biosynthesis of amino acids, glucosinolate biosynthesis and valine, leucine and isoleucine biosynthesis were the top-5 KEGG enriched pathways when comparing the CG and DS metabolome (M_CG/DS). Comparing DS with COS (M_DS/COS) showed that purine metabolism and phenylalanine metabolism were enriched. Combined transcriptome and metabolome analysis revealed that pyruvate and phenylalanine metabolism were KEGG-enriched pathways for CG/DS and DS/COS, respectively. For pyruvate metabolism, 87 DEGs (47 up-regulated and 40 down-regulated) and five SDMs (1 up-regulated and 4 down-regulated) were enriched. Pyruvate was closely related with 14 DEGs (|r| > 0.99) after Pearson's correlation analysis, and only 1 DEG (Sspon.02G0043670-1B) was positively correlated. For phenylalanine metabolism, 13 DEGs (7 up-regulated and 6 down-regulated) and 6 SDMs (1 up-regulated and 5 down-regulated) were identified. Five PAL genes were closely related with 6 SDMs through Pearson's correlation analysis, and the novel.31257 gene had significantly up-regulated expression. Collectively, our results showed that DS has significant adverse effects on the physiology, transcriptome, and metabolome of sugarcane, particularly genes involved in photosynthesis. We further show that COS treatment can alleviate these negative effects.

First Report of Pantoea stewartii subsp. stewartii Causing Bacterial Leaf Wilt of Sugarcane in China.

The pathogen of Pantoea stewartii subsp. stewartii (Pss) that is the causal agent of Stewart's bacterial wilt of corn also infects numerous experimental hosts of graminaceous plants (Pepper et al., 1967; Wang et al., 2012). However, little is known about this pathogen naturally infecting sugarcane. In 2017, we observed some sugarcane cultivars showing leaf blade bleaching at the disease initiation stage, which further resulted in development of blight and necrotic lesions (Figure 1-A and -B) in Zhanjiang, Guangdong province of China. To diagnose this putative disease, five symptomatic leaf samples were collected from different sugarcane cultivars. The Pss was found to infect these samples using the nested PCR with Pss-specific outer primers PS1/PS4 and inner primers Ps2r/Ps3r that targeted at the 16S rRNA gene of this pathogen (Wang et al., 2009). The expected 262-bp fragments from positive samples were amplified, cloned, and sequenced (GenBank accession no. MW015795-MW015799). BLASTN analysis revealed that these isolates had more than 99.5% nucleotide identify (222 bp out of 262 bp) with each other and with Pss strains (ATCC 8199 and DC283) as well as P. stewartii subsp. indologenes strains (SR2-12 and LMG 2632) after sequences were trimmed at the 5'- and 3'-terminal of inner primer sequences. In addition, these leaf samples were surface-sterilized with 75% alcohol followed by macerated and chopped in sterile water. Upon plating on solid NA medium at 28 °C for 24-36 h, the bacterial colonies exhibited yellow color with circular, convex, smooth and translucent edges (Figure 1-C). Straight rods and non-encapsulated cells were detected under transmission electron microscopy (Figure 1-D). Moreover, an identical colony termed as PSCN1 was isolated from sugarcane cultivar YZ08-1095 and was further confirmed by the PCR with a universal primer pair 63F (5'-CAGGCCTAACACATGCAAGTC-3') and 1387R (5'-GGGCGGWGTGTACAAGGC-3') that targeted at bacterial 16S rRNA gene (Marchesi et al., 1998). A 1362-bp DNA fragment sequence was obtained from PSCN1 strain and deposited on GenBank library (accession no. MW015767). Sequence analysis showed that PSCN1 shared 99.9-100% nucleotide identity (1315 bp out of 1362 bp) with the two reference strains of Pss (ATCC 8199 and DC283) after sequences were trimmed at the 5'- and 3'-terminal of primer sequences. According to Koch's postulates, pathogenicity test was carried out on YZ08-1095 plants with 3-5 fully developed leaf inoculated with the suspended cells (108 cells/ml) of PSCN1 strain by cutting the one-third of leaves before spraying with a suspension. Control plants were mock-inoculated with serial liquid nutrition agar medium. Two independent experiments were performed for pathogenicity assay and more than 28 plants of YZ08-1095 were used in each treatment. Plants were cultured in a growth chamber at 28 °C and 60% humidity under a 16 h light/8 h dark photoperiod. Leaves inoculated by the PSCN1 initially showed bleached, blight and wilting symptoms on leaf edges at seven days post-inoculation (dpi) (Figure 1-E and -F), which were similar to those symptoms observed in the fields. Control plants remained asymptomatic (Figure 1-G). The average incidence of diseased plant was 51.9% at 21 dpi. The bacteria were subsequently re-isolated from diseased leaves, and yielded colonies were completely identical to the PSCN1. Taken together, our data provides the valuable information for diagnosis and controlling this disease in sugarcane.

Energy Partitioning and Latent Heat Flux Driving Factors of the CAM Plant Pineapple (Ananas comosus (L.) Merril) Grown in the South Subtropical China.

Elucidation of different vegetation energy partitioning and environmental control factors at the agro-ecosystem levels is critical for better understanding and scientific management of farmland. Pineapple (Ananas comosus (L.) Merril) is a tropical plant widely cultivated in the southern subtropical region of China; however, the energy partitioning of crassulacean acid metabolism (CAM) plants like pineapple and their interactions with the environment remain not well understood. In this study, we investigated the energy partitioning patterns of pineapple fields and latent heat flux (LET) response to environmental factors using the Bowen ratio energy balance system and meteorological observation field data. The results showed that the CAM plant pineapple energy partitioning was significantly different from the common C3 and C4 crops during the study period, which was mainly attributed to the complex interactions between CAM plant transpiration and the environment. Specifically, sensible heat flux was the main component of net radiation (Rn), followed by the LET, accounting for 65.0% and 30.8% of the Rn, respectively. Soil heat flux accounts for a very small fraction (4.2%). The mean values of the Bowen ratio were 2.09 and 1.41 for sunny and cloudy days during the daytime and 0.74 and 0.46 at night, respectively. LET is a key factor in responding to crop growth status and agricultural water management, and the path analysis indicates that its variation is mainly influenced directly by Rn with a direct path coefficient of 0.94 on sunny days, followed by vapor pressure deficit, air temperature and relative humidity, which indirectly affect LET through the Rn pathway, whereas soil moisture and wind speed have a low effect on LET. On cloudy days, the effect of Rn on LET was overwhelmingly dominant, with a direct path coefficient of 0.91. The direct path coefficients of the remaining factors on LET were very small and negative. Overall, this study is an important reference for enhancing the impact of pineapple as well as CAM plants on the surface energy balance and regional climate.

Evaluation of a Sugarcane (Saccharum spp.) Hybrid F1 Population Phenotypic Diversity and Construction of a Rapid Sucrose Yield Estimation Model for Breeding.

Sugarcane is the major sugar-producing crop worldwide, and hybrid F1 populations are the primary populations used in breeding. Challenged by the sugarcane genome's complexity and the sucrose yield's quantitative nature, phenotypic selection is still the most commonly used approach for high-sucrose yield sugarcane breeding. In this study, a hybrid F1 population containing 135 hybrids was constructed and evaluated for 11 traits (sucrose yield (SY) and its related traits) in a randomized complete-block design during two consecutive growing seasons. The results revealed that all the traits exhibited distinct variation, with the coefficient of variation (CV) ranging from 0.09 to 0.35, the Shannon-Wiener diversity index (H') ranging between 2.64 and 2.98, and the broad-sense heritability ranging from 0.75 to 0.84. Correlation analysis revealed complex correlations between the traits, with 30 trait pairs being significantly correlated. Eight traits, including stalk number (SN), stalk diameter (SD), internode length (IL), stalk height (SH), stalk weight (SW), Brix (B), sucrose content (SC), and yield (Y), were significantly positively correlated with sucrose yield (SY). Cluster analysis based on the 11 traits divided the 135 F1 hybrids into three groups, with 55 hybrids in Group I, 69 hybrids in Group II, and 11 hybrids in Group III. The principal component analysis indicated that the values of the first four major components' vectors were greater than 1 and the cumulative contribution rate reached 80.93%. Based on the main component values of all samples, 24 F1 genotypes had greater values than the high-yielding parent 'ROC22' and were selected for the next breeding stage. A rapid sucrose yield estimation equation was established using four easily measured sucrose yield-related traits through multivariable linear stepwise regression. The model was subsequently confirmed using 26 sugarcane cultivars and 24 F1 hybrids. This study concludes that the sugarcane F1 population holds great genetic diversity in sucrose yield-related traits. The sucrose yield estimation model, ySY=2.01xSN+8.32xSD+0.79xB+3.44xSH-47.64, can aid to breed sugarcane varieties with high sucrose yield.

Simulation of Photosynthetic Quantum Efficiency and Energy Distribution Analysis Reveals Differential Drought Response Strategies in Two (Drought-Resistant and -Susceptible) Sugarcane Cultivars.

Selections of drought-tolerant cultivars and drought-stress diagnosis are important for sugarcane production under seasonal drought, which becomes a crucial factor causing sugarcane yield reduction. The main objective of this study was to investigate the differential drought-response strategies of drought-resistant ('ROC22') and -susceptible ('ROC16') sugarcane cultivars via photosynthetic quantum efficiency (Φ) simulation and analyze photosystem energy distribution. Five experiments were conducted to measure chlorophyll fluorescence parameters under different photothermal and natural drought conditions. The response model of Φ to photosynthetically active radiation (PAR), temperature (T), and the relative water content of the substrate (rSWC) was established for both cultivars. The results showed that the decreasing rate of Φ was higher at lower temperatures than at higher temperatures, with increasing PAR under well-watered conditions. The drought-stress indexes (εD) of both cultivars increased after rSWC decreased to the critical values of 40% and 29% for 'ROC22' and 'ROC16', respectively, indicating that the photosystem of 'ROC22' reacted more quickly than that of 'ROC16' to water deficit. An earlier response and higher capability of nonphotochemical quenching (NPQ) accompanied the slower and slighter increments of the yield for other energy losses (ΦNO) for 'ROC22' (at day5, with a rSWC of 40%) compared with 'ROC16' (at day3, with a rSWC of 56%), indicating that a rapid decrease in water consumption and an increase in energy dissipation involved in delaying the photosystem injury could contribute to drought tolerance for sugarcane. In addition, the rSWC of 'ROC16' was lower than that of 'ROC22' throughout the drought treatment, suggesting that high water consumption might be adverse to drought tolerance of sugarcane. This model could be applied for drought-tolerance assessment or drought-stress diagnosis for sugarcane cultivars.

Homogeneous isolation of nanocellulose from sugarcane bagasse by high pressure homogenization.

Nanocellulose from sugarcane bagasse was isolated by high pressure homogenization in a homogeneous media. Pretreatment with an ionic liquid (1-butyl-3-methylimidazolium chloride ([Bmim]Cl)) was initially involved to dissolve the bagasse cellulose. Subsequently, the homogeneous solution was passed through a high pressure homogenizer without any clogging. The nanocellulose was obtained at 80 MPa for 30 cycles with recovery of 90% under the optimum refining condition. Nanocellulose had been characterized by Fourier transformed infrared spectra, X-ray diffraction, thermogravimetric analysis, rheological measurements and transmission electron microscopy. The results showed that nanocellulose was 10-20 nm in diameter, and presented lower thermal stability and crystallinity than the original cellulose. The developed nanocellulose would be a very versatile renewable material.

[Preliminary clinical study of application of cyntridini triphosphatis and dexamethasone in treatment of sudden deafness].

OBJECTIVE: To investigate the effect of application of cytridini triphosphatis (CTP) combined with dexamethasone on sudden deafness. METHODS: Two-hundred and seven patients (312 ears) with sudden deafness were randomly divided into CTP group (n = 159 ears) and control group (n = 153 ears). Besides intravenous administration of CTP, another treatments in CTP group were the same as that in control group including hyperbaric oxygen, vasodilator, energy preparation such as ATP, CoA and dexamethasone (DXM). RESULTS: The hearing threshold was (75.56 +/- 30.24) dB HL in CTP group and (72.50 +/- 40.50)dB HL in control group (P > 0.05) before treatment. The average value of the hearing threshold after treatment was decreased by (50.08 +/- 21.47) dB HL in CTP group and (19.45 +/- 19.12) dB in control group(P < 0.05), respectively. CONCLUSION: The application of CTP combined with DXM can significantly improve the effect of treatment on sudden deafness and CTP may enhance the recovery rate in the patients with sudden deafness, which was much higher than that treated by the traditional methods. However, the mechanism of the effect of CTP coupled with DXM is still unknown and further study is necessary.