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Sugarcane white leaf (SCWL) is a phytoplasma disease that has spread rapidly in Yunnan province, China, in recent years. It is now the main barrier to the development of the sugarcane industry in this region. Herein, we explored the insect vector and their dominant population of SCWL phytoplasma at Gengma in Lincang city in 2019, where the occurrence of SCWL is the most serious in Yunnan Province. Two leafhopper species, Tettigoniella viridis (Linnaeus) and Clovia conifer Walker, were tested positive for SCWL phytoplasma by using nested PCR, indicating that they are maybe the potential insect vectors of SCWL phytoplasma. Additionally, nymphs of T. viridis were tested strongly positive, and they were preliminarily identified as the dominant population. Since the main transmission medias of SCWL phytoplasma appears to be SCWL-infected seedcane and the leafhopper insect vectors, establishing a disease-free and healthy seedcane breeding base and eliminating the leafhopper insect vectors in sugarcane planting areas may prevent or control the spread of SCWL and reduce the natural transmission rate in the field. The results expand our theoretical and technical knowledge of sugarcane phytoplasma disease and provide a reference for formulating comprehensive control of SCWL.

Brown stripe is a major fungal disease damaging the leaves of sugarcane. Breeding and cultivating resistant varieties is the most economical and effective method of controlling the disease. The paper reports on the evaluation of sugarcane brown stripe resistance in novel varieties bred by the China Sugarcane System and major varieties under cultivation in the sugarcane cultivating region. Field resistance against brown stripe was identified in 60 novel varieties and in 31 major varieties under cultivation in Lincang, Puer, and Yuxi, in Yunnan Province, and in Yizhou, Guangxi Province, where the incidence and severity of brown stripe is particularly high. Field surveys showed that 32 (53.3%) of the 60 novel varieties were highly resistant to moderately resistant, and 28 (46.7%) were susceptible to highly susceptible. In addition 21 (67.7%) out of the 31 major cultivated varieties were highly resistant to moderately resistant, and 10 (32.3%) were susceptible to highly susceptible. We recommend the intensifying of efforts to eliminate the major varieties that are susceptible, and promotion of the adoption of elite resistant varieties in wet and rainy sugarcane cultivating regions with high incidence and severity of sugarcane brown stripe disease, to effectively manage the brown stripe epidemic.

We describe a new α-globin chain variant in a Chinese subject. This novel variant, with a Val→Met substitution at codon 93 of the α-globin chain, has been named Hb Qingcheng (HBA1: c.280G>A) for where the proband was born. A woman with somatic mosaicism for Hb Qingcheng presented with the phenotype of mild α-thalassemia (α-thal).

Sugarcane brown rust, caused by Puccinia melanocephala, is one of the main diseases of sugarcane in China. The identification and discovery of new resistance genes have important theoretical and practical significance for preventing outbreaks of brown rust and ensuring the sustainable production of sugarcane. To screen for polymorphic simple-sequence repeat (SSR) molecular markers for localization of brown rust resistance genes, we used two populations that are suitable for genetic linkage map construction and mapping of new resistance genes to construct resistant and susceptible genetic pools. We then screened 449 pairs of primers to identify polymorphic SSR markers in the parental lines and the resistant/susceptible genetic pools. The results showed that 25 pairs of primers directed amplification of polymorphic DNA fragments between the parents of the cross combination ‘Yuetang 03-393’ × ‘ROC 24’, and 16 pairs of primers amplified polymorphic fragments between the parents of the cross combination ‘Liucheng 03-1137’ × ‘Dezhe 93-88’. Four pairs of primers (SMC236CG, SCESSR0928, SCESSR0636 and SCESSR2551) amplified polymorphic DNA fragments between the parental lines and the resistant/susceptible genetic pools in ‘Yuetang 03-393’ × ‘ROC 24’. The results of this study will establish a solid foundation for the mapping of new brown rust resistance genes, genetic linkage map construction and the development of closely-associated molecular markers in sugarcane.

The aim of this study was to assess brown rust resistance of new sugarcane varieties bred in China and main cultivated varieties in sugarcane planting area, and screen the elite new brown rust resistant varieties for popularization and application in production. In total, 60 new varieties in the national regional test of new sugarcane varieties were tested in Kaiyuan and Lincang, and 34 main cultivated varieties were studied in Lincang, Puer, and Yuxi, Yunnan province, and Yizhou, Guangxi province, China, where the incidence of brown rust was particularly high. The resistance of these sugarcane varieties to brown rust was investigated under natural inoculation and molecular marker-assisted identification was used to detect the brown rust resistance gene Bru1 . The results of field survey showed that 66 (70.21%) of the 94 new and main cultivated varieties were highly resistant to moderately resistant, and 28 (29.79%) were susceptible to highly susceptible. Molecular detection indicated that Bru1 gene was found among 54 (57.45%) of the 94 new and main cultivated varieties. Some main cultivated varieties that were currently planted across large areas such as Guitang 29, Guitang 44, Dezhe 03-83, Liucheng 03-1137, Yuetang 60, and Guitang 46 were highly susceptible to brown rust, and 31 new varieties such as Yuegan 48, Funong 09-2201, Guitang 08-120, Liucheng 09-15, Zhongzhe 1, Yunzhe 08-1609, Yunrui 10-187, and Zhongtang 1201 were resistant. Therefore, in the sugarcane planting areas with high incidence of brown rust and wet and rainy climates, more effort should be eliminated the main susceptible varieties and promoted the application of new resistant varieties. This will help to achieve a reasonable distribution of varieties, fundamentally control the outbreak of disease in sugarcane planting areas, and provide security for the high-quality development of sugarcane industry in China in the future.

To test high cadmium (Cd) concentration may not be high in health risk when considering Cd bioavailability, we assessed variation of Cd relative bioavailability (RBA, relative to CdCl2) using a mouse assay for 14 vegetables of water spinach, amaranth, and pakchoi. Cadmium concentration varied from 0.13 ± 0.01–0.37 ± 0.00 μg g–1 fw. Cadmium-RBA also varied significantly from 22.9 ± 2.12–77.2 ± 4.46%, however, the variation was overall opposite to that of Cd concentration, as indicated by a strong negative correlation between Cd-RBA and Cd concentration (R2 = 0.43). Based on both Cd concentration and bioavailability, the identified high-Cd pakchoi variety resulted in significantly lower Cd intake than the high-Cd varieties of water spinach and amaranth (4.74 ± 0.05 vs. 10.1 ± 0.54 and 8.03 ± 0.04 μg kg–1 bw week–1) due to significantly lower Cd-RBA (22.9 ± 2.12 vs. 77.2 ± 4.46 and 51.3 ± 2.93%). The lower Cd-RBA in pakchoi was due to its significantly higher Ca and lower phytate concentrations, which facilitated the role of Ca in inhibiting intestinal Cd absorption. This was ascertained by observation of decreased Cd-RBA (90.5 ± 12.0% to 63.5 ± 5.53%) for a water spinach when elevating its Ca concentration by 30% with foliar Ca application. Our results suggest that to assess food Cd risk, both total Cd and Cd bioavailability should be considered. Refereed/Peer-reviewed

Pokkah boeng is an epidemic fungal disease that greatly affects the development of the sugarcane industry in China. Resistance to pokkah boeng disease differs among sugarcane varieties. Breeding and planting resistant cultivars is the most economic and effective measure for controlling this disease. In this study, new sugarcane varieties bred by sugarcane research units in China and the major varieties cultivated in various sugarcane areas, were screened to find new elite pokkah boeng-resistant varieties for application in sugarcane production. Sixty new varieties resistance to pokkah boeng disease were evaluated in regionalized experiments under field conditions. Meanwhile, the resistance of the 31 main cultivated varieties was analyzed under field conditions in Lincang, Puer, and Yuxi, Yunnan Province, and in Yizhou, Guangxi Province, where the incidence of pokkah boeng is particularly high. Field surveys showed that 35 (58.3%) of the 60 new sugarcane varieties were highly resistant to moderately resistant, and 25 (41.7%) were susceptible to highly susceptible. Of the 31 main cultivated varieties, 15 (48.4%) were highly resistant to moderately resistant, and 16 (51.6%) were susceptible to highly susceptible. The results suggested that, in wetter and rainy sugarcane cultivation areas with high incidence of pokkah boeng disease, it is necessary to increase efforts to eliminate the susceptible major cultivated varieties and promote the cultivation of novel resistant varieties. This will help to achieve a reasonable distribution of varieties, fundamentally control the outbreak of pokkah boeng disease, and facilitate the high-quality development of the sugarcane industry in China.

The aim of this study was to assess brown rust resistance in 60 new sugarcane varieties bred by the China Sugarcane System in recent years and in 34 major varieties cultivated in sugarcane growing areas in China. The resistance of these sugarcane varieties to brown rust was investigated in the field, and molecular markers were used to detect the brown rust-resistance gene Bru1. The results of the field survey showed that 66 (70.21%) of the 94 sugarcane varieties were highly resistant to moderately resistant, and 28 (29.79%) were susceptible to highly susceptible. The Bru1 gene was detected in 54 (57.45%) of the 94 sugarcane varieties. Seven highly resistant varieties and five resistant varieties did not carry Bru1, suggesting that they carry other genes associated with brown rust resistance. This study provides a scientific basis and identifies disease-resistant germplasm for selection of plants for sugarcane production.

BACKGROUND Acute kidney injury (AKI) is a common and multifactorial complication after liver transplantation (LT). Myoglobin (Mb) which can be served as O2 storage and delivery depot is present in muscles and cardiac myocytes. Previous studies had shown the close relationship between Mb and AKI. But there is a lack of clinical studies for Mb with the risk of AKI due to LT. This study was performed to determine the association between the serum level of Mb and incidence of AKI in patients underwent LT. METHODS The clinical data of 140 consecutive adult patients who underwent LT at our center from June 2018 to August 2020 were analyzed in this study. One hundred and fifteen patients met the inclusion criteria. The performances of postoperative laboratory variables (including serum Mb) were evaluated. The outcomes after LT, including the duration of intensive care unit (ICU) stay, hospital stay and 28-day mortality, were also measured. RESULTS We divided 115 patients into AKI group (n=44) and non-AKI group (n=71). Serum Mb on postoperative day 0 (POD0) was significantly higher in AKI group than those in non-AKI group (P

Ratoon stunting disease (RSD) and brown rust are major plant diseases that disrupt sugarcane production. Identifying disease-resistant parental lines offers an efficient strategy for controlling these diseases in sugarcane. The aim of this study was to clarify the occurrence of RSD and the distribution of the major brown rust resistance gene Bru1 among commonly used sugarcane breeding parents in China. We used PCR assays for the molecular detection of RSD and Bru1 in 255 sugarcane breeding parents collected from the National Germplasm Repository of Sugarcane of China (2019–2020). RSD was detected in 68.6% of the parental lines, indicating that commonly used sugarcane breeding parents in China are severely infected by RSD. However, 80 of the tested lines were RSD negative, indicating these parental lines had good resistance to RSD. The RSD incidence rate varied between different parental line series: VMC had the lowest RSD frequency (40.0%) and Guitang had the highest frequency (84.6%), indicating that the VMC series had good resistance levels to RSD and that a high proportion of parental lines were RSD resistant. Bru1 was present in 87 of the 255 tested parental lines, with greatest frequency detected in the ROC series (71.4%) and lowest frequency detected in the CP series (21.6%). These results indicate that Bru1 is a major source of brown rust resistance in commonly used sugarcane breeding parents in China, with important implications for brown rust resistance breeding programs in sugarcane. Overall, these results provide a reference for selecting appropriate breeding lines that are resistant to RSD and brown rust, offering guidance for effective disease control in sugarcane.

To improve the proteolytic stability of the lipase LIP2 from Yarrowia lipolytica, the peptide bonds susceptible to trypsin in LIP2 were analyzed by tandem mass spectrometry and redesigned by site-directed mutagenesis. Different variants of the enzyme were expressed in Pichia pastoris GS115 and their biochemical properties were subsequently investigated. Although most of the variants were still cleaved by trypsin, some of them did show an evident increase of resistance against proteolytic degradation. The most stable mutant was LIP2-C5, in which five trypsin-cleavage sites were replaced by non-preferred amino acids. Upon incubation with human trypsin for 80 min at 37°C, the mutant LIP2-C5 was found to retain >70% of its initial activity, compared to only 10% for the wild-type.

Sugarcane brown rust induced by Puccinia melanocephala is an important global disease. Exploring novel resistance genes and breeding varieties with durable resistance is the most economical and effective way of controlling brown rust. The establishment of complete and high-density molecular genetics maps is a key prerequisite for the discovery and location of sugarcane brown rust resistance genes. To obtain more polymorphic simple sequence repeat (SSR) markers for the construction of genetic linkage maps, six highly resistant varieties and six highly susceptible varieties were used as parents to screen the primers with clear bands, obvious polymorphism, and good repeatability. The highest numbers of polymorphic primers were obtained from the parental group of Liucheng 03-1137 × Dezhe 93-88 (52.38%). Ten pairs of primers including mSSCIR34 exhibited high polymorphism in the parental group of Liucheng 03-1137 × Dezhe 93-88. Here, we recommend ten SSR pairs of polymorphic primers including mSSCIR34 to use in the mapping populations of Liucheng 03-1137 × Dezhe 93-88. The polymorphic primers could facilitate the construction of genetic maps and lay an appropriate foundation for the localization of brown rust resistance genes and development of molecular genetics linkage markers.

We assessed inheritance of resistance to sugarcane brown rust (Puccinia melanocephala) in selfing F1 populations of wild sugarcane germplasm Erianthus rockii ‘Yundian 95-19’ and E. rockii ‘Yundian 95-20’. We tested parent and selfing F1 individuals for the brown rust resistance gene, Bru1, that has been shown to confer resistance to brown rust in sugarcane. The Bru1 gene was not detected in E. rockii ‘Yundian 95-19’, E. rockii ‘Yundian 95-20’ or their selfing F1 individuals, and we found there was segregation of resistance in the two selfing F1 populations (segregation ratio: 3:1). The results confirmed resistance in E. rockii ‘Yundian 95-19’ and E. rockii ‘Yundian 95-20’ to sugarcane brown rust is controlled by a novel, single dominant gene.

Sugarcane leaf scald caused by Xanthomonas albilineans (Ashby) Dowson is an extremely destructive disease. In 2017, suspected symptoms of sugarcane leaf scald was observed, in Baoshan, Mengding and Jinping, Yunnan Province, China. The bacteria was isolated from the symptomatic stalks and was subsequently identified as the causal agent using Koch’s postulates. BLASTN analysis showed that the amplified nucleotide sequences of 16S rRNA, gyrB and rpoD genes from all isolates shared 100% sequence identity with the nucleotide sequences of corresponding genes of X. albilineans strain GPE PC73 (Acc no. FP565176). Based on symptom diagnosis in the field, colony morphology, validation of Koch’s postulates and molecular identification of 16S rRNA, gyrB and rpoD genes, the disease was confirmed as sugarcane leaf scald caused by X. albilineans. Partial 16S rRNA, gyrB and rpoD gene sequence analysis of 36 X. albilineans strains from Yunnan and Guangxi provinces, China, was determined. The results revealed that the gyrB gene was the most sensitive in differentiating X. albilineans strains from Yunnan and Guangxi provinces.

Background: Spinal cord injury (SCI) is a worldwide medical concern. This study aimed to elucidate the mechanism underlying the protective effect of hyperbaric oxygen (HBO) against SCI-induced neurologic defects in rats via exploring the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis and expression of brain-derived neurotrophic factor (BDNF). Methods: An acute SCI rat model was established in Sprague-Dawley rats using the Allen method. Sixty rats were divided into four groups (n= 15 in each group): sham-operated, SCI, SCI treated with HBO (SCI+ HBO), and SCI treated with both HBO and AMD3100 (an antagonist of CXCR4; SCI+ HBO+ AMD) groups. The rats were treated with HBO twice a day for 3 days and thereafter once a day after the surgery for up to 28 days. Following the surgery, neurologic assessments were performed with the Basso-Bettie-Bresnahan (BBB) scoring system on postoperative day (POD) 7, 14, 21, and 28. Spinal cord tissues were harvested to assess the expression of SDF-1, CXCR4, and BDNF at mRNA and protein levels, using quantitative real-time polymerase chain reaction, Western blot analysis, and histopathologic analysis. Results: HBO treatment recovered SCI-induced descent of BBB scores on POD 14, (1.25 ± 0.75 vs. 1.03 ± 0.66, P < 0.05), 21 (5.27 ± 0.89 vs. 2.56 ± 1.24, P < 0.05), and 28 (11.35 ± 0.56 vs. 4.23 ± 1.20, P < 0.05) compared with the SCI group. Significant differences were found in the mRNA levels of SDF-1 (mRNA: day 21, SCI+ HBO vs. SCI+ HBO+ AMD, 2.89 ± 1.60 vs. 1.56 ± 0.98, P < 0.05), CXCR4 (mRNA: day 7, SCI+ HBO vs. SCI, 2.99 ± 1.60 vs.1.31 ± 0.98, P < 0.05; day 14, SCI+ HBO vs. SCI+ HBO+ AMD, 4.18 ± 1.60 vs. 0.80 ± 0.34, P < 0.05; day 21, SCI+ HBO vs. SCI, 2.10 ± 1.01 vs.1.15 ± 0.03, P < 0.05), and BDNF (mRNA: day 7, SCI+ HBO vs. SCI, 3.04 ± 0.41 vs. 2.75 ± 0.31, P < 0.05; day 14, SCI+ HBO vs. SCI, 3.88 ± 1.59 vs. 1.11 ± 0.40, P < 0.05), indicating the involvement of SDF-1/CXCR4 axis in the protective effect of HBO. Conclusions: HBO might promote the recovery of neurologic function after SCI in rats via activating the SDF-1/CXCR4 axis and promoting BDNF expression. Key words: Brain-derived neurotrophic factor; CXC chemokine receptor 4; Hyperbaric oxygen; Neurotrophic; Stromal cell-derived factor-1; Spinal cord injury

Caragana korshinskii Kom., an arbuscular legume with important economic and ecological value in feed, processing industry, and environmental protection, also has great tolerance potential to abiotic stress conditions. An AP2 domain-containing gene was isolated from the suppression subtractive hybridization library of C. korshinskii under drought stress. In addition, the isolated gene was also found to be responsive to cold and ABA treatment. Phylogenetic analysis indicates that the deduced protein belongs to the DREB A-1 subfamily and is designated as CkDREB1. Overexpression of CkDREB1 in Arabidopsis thaliana (L.) Heynh increased drought and cold tolerance compared with the wild type. The drought responsive genes RD29A, RD29B, KIN1, and KIN2, as well as cold-responsive marker genes COR15A and COR47, were also highly induced in the overexpression lines under drought and cold conditions. These results should shed light on our understanding on the mechanisms of abiotic resistance of C. korshinskii.

Sugarcane (Saccharum officinarum L.) is the main sugar crop in China. Yunnan is the second largest sugarcane production province in China. In December 2018, leaf blight was first observed on almost every leaf of sugarcane on 'Huanan 54-11', 'Baimei' and 'Chongan' in Kaiyuan (103°27' E, 23°72' N), Yunnan. In October 2019, during our survey in the field in Lingcang (100°08' E, 23°88' N), Yunnan, this disease was also observed on 'ROC 25'. Symptoms of the disease initially appeared as wilted, which seemed to be cause by water stress. As the disease progressed, irregular straw-yellow and blighted lesion ran throughout the leaf lamina from leaf tip to entire leaf sheath, many small black conidia formed in the dead leaf tissue under humid conditions. Symptomatic leaf tissues were surface-sterilized with 70% ethanol for 30 s, 0.1% HgCl2 for 1 min, and rinsed with sterilized water three times, air dried on sterile filter paper, and plated on potato dextrose agar (PDA). Six isolates were obtained from six symptomatic leaf samples and were transferred onto potato carrot agar (PCA). Colonies on PDA were white with loose aerial hyphae at first, then turned to dark olive or dark. Colonies on PCA were grayish with sparse hyphae, then turned to dark gray. Conidiophores were brown, simple or branched, and produced numerous conidia in short chains. Conidia (n = 50) were obclavate to obpyriform or ellipsoid, brown to dark brown, with a cylindrical short beak at the tip (2.3 to 17.3 µm in length), and 15.3 to 46.6 μm × 4.2 to 17.9 μm, 2 to 7 transverse septa and 0 to 3 longitudinal septa. Morphologically, the isolates were identified as Alternaria tenuissima (Simmons 2007). Two representative isolates C4 and C5 were selected for molecular identification. The internal transcribed spacers (ITS), Histone 3 genes and plasma membrane ATPase were amplified with primer pairs ITS1/ITS4, H3-1a/H3-1b and ATPDF1/ATPDR1, respectively (Glass et al. 1995; Lawrence et al. 2013). The sequences were deposited in GenBank (ITS, MT679707-MT679708; Histone 3, MT710929-MT710930; ATPase, MT833928-MT833929). BLAST searches showed ≥99% nucleotide identity to the sequence of A. tenuissima (ITS, 100% to MN822571; Histone 3, 100% to MN481955; ATPase, 99% to JQ671875, 100% to MH492703, respectively). Thus, the fungus was identified as A. tenuissima based on morphological and molecular characteristics. For pathogenicity tests, five healthy 2-month-old potted sugarcane leaves were wounded with one sterile needle and inoculated with 20 μl of suspension of 106 conidia/ mL, and five plants were inoculated with distilled water as the controls. Plants were placed in a greenhouse at 25 to 35°C. After two months, the leaf wound inoculated with the putative pathogen displayed blighted as those observed in the field whereas the controls remained symptomless. The fungus was reisolated from symptomatic leaves with the same morphological and molecular traits as the original isolates. The fungus was not isolated from the control plants. Pathogenicity tests were repeated two times. A. tenuissima causing leaf blight on barley in China was reported in 2008 (Luo et al. 2008). Leaf spot disease of sugarcane caused by A. tenuis has been recorded in Maharashtra (Patil et al. 1974). To our knowledge, this is the first report on A. tenuissima affecting leaf blight on sugarcane in Yunnan Province, China. Identification of the causes of the disease is important to develop effective disease management strategies. The author(s) declare no conflict of interest. Funding: This research was supported by Sugar Crop Research System (CARS-170303), the Yunling Industry and Technology Leading Talent Training Program "Prevention and Control of Sugarcane Pests" (2018LJRC56), and the Yunnan Province Agriculture Research System. References: Glass, N. L., et al. 1995. Appl. Environ. Microbiol. 61:1323. Lawrence, D. P., et al. 2013. Mycologia 105:530. Luo, Z., et al. 2008. Acta Phytophy. Sin. 35(5): 469-470. Patil, A.O., et al. 1974. Res. J. Mahatma Phule Agric. Univ. 5(2): 122-123. Simmons, E. G. 2007. Alternaria: An Identification Manual. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands. Caption for supplementary Figure 1 Supplementary Figure S1. Disease symptoms of sugarcane leaf blight disease and morphological characteristics of Alternaria tenuissima. (A) Typical straw-yellow and blighted lesions on naturally-infected leaves of sugarcane; (B) Infected symptoms on wounded leaves of sugarcane two months after artificial infection with A. tenuissima; (C) Colony of A. tenuissima on PDA; (D) Colony of A. tenuissima on PCA; and (E-F) Sporulation and conidia of A. tenuissima on PCA. (Scale bars = 100 μm; 20 μm).

Sugarcane white leaf (SCWL) is a devastating sugarcane (Saccharum officinarum) disease caused by a 16SrXI group phytoplasma, which is extremely harmful to sugarcane production. To determine the occurrence of SCWL in different varieties in 2018, we conducted a field survey and performed nested PCR detection of SCWL phytoplasma in cane-planting areas of Mangweng and Hepai in Gengma, Yunnan province, which are the areas most severely affected by SCWL in China. The results of the field survey showed that the symptomatic incidence of SCWL differed among varieties. The mean symptomatic incidence of SCWL on variety Yuetang60 was the highest (73.50%), and it was the lowest on Liucheng05-136 (13.67%). Using nested PCR, the SCWL phytoplasma was detected in symptomatic plants of all varieties more than 90% of the time; the SCWL phytoplasma was detected in 91 and 97% of symptomatic plants of Yingyu91-59 and Liucheng05-136 varieties, respectively. The SCWL phytoplasma was detected by PCR in 82% of the asymptomatic plant samples. The results of this study showed that field survey based on white leaf symptoms did not accurately reflect the actual occurrence of the SCWL phytoplasma.

The biosynthesis of the valuable antibiotic enduracidin by Streptomyces fungicidicus TXX3120 is a complex multistep process. To identify the rate-limiting step of the entire biosynthetic process, we carried out a deep RNA sequencing towards the mycelia of TXX3120 at different fermentation stages. Comparative RNA-seq analysis indicated that the expression level of the endC gene during the enduracidin production phase was evidently lower than that of the other relevant genes to enduracidin biosynthesis. This result was further confirmed by quantitative RT-PCR, and the giant non-ribosomal peptide synthase (NRPS) encoded by endC was predicated to be the rate-limiting enzyme in enduracidin biosynthesis. To increase the expression of endC during the enduracidin production phase, a reporter-based selection system was developed by genetically replacing the initial part of the endC gene with a thiostrepton resistance gene (tsr), which will then act as a selectable marker to report the expression level of the rate-limiting gene endC, thereby facilitating the selection of enduracidin-overproducing mutants following random mutagenesis. After one round of mutagenesis, thiostrepton resistance selection, and restoration of the endC gene, three mutant strains with improved endC expression levels were obtained. Their highest enduracidin titers reached 9780.54, 9272.46, and 8849.06 U/mL, respectively representing 2.31-, 2.19-, and 2.09-fold of the initial industrial strain TXX3120. Our research provides a useful strategy for the rational breeding of industrial strains that synthesize complex natural products.

Sugarcane mosaic disease is one of the most serious and prevalent viral diseases of sugarcane in China. Resistant varieties are the most economical and effective measures for controlling this disease. Resistance against Sugarcane streak mosaic virus (SCSMV) and Sorghum mosaic virus (SrMV) in 41 excellent sugarcane innovation germplasms and their parents (Saccharum officinarum L. “Ludashi” × Erianthus rockii Keng “Yundian 95-19”) was checked by using a stalk inoculation and RT-PCR detection during 2015 and 2016. Results indicated that among the 41 excellent sugarcane innovation germplasms and parents, 23 were highly (Grade 1) to moderately resistant (Grade 3) to SCSMV, meanwhile, 31 were highly (Grade 1) to moderately resistant (Grade 3) to SrMV. Ten germplasms were highly resistant (Grade 1) to resistant (Grade 2) to both SCSMV and SrMV. Among these, six germplasms (Yun 09-604, Yun 09-607, Yun 09-619, Yun 09-633, Yun 09-656, Yundian 95-19) were highly resistant (Grade 1) to both SCSMV and SrMV, accounting for 13.95% of the total germplasm materials. These results may provide resistance resource available for the breeding program of sugarcane cultivars against the major vrius associated with mosaic disease in China.

Field resistance against sugarcane thrips (Fulmekiola serratus) in 21 sugarcane varieties was tested following inoculation with a natural population. The results revealed differences in pest incidence levels in different varieties and between different leaf positions in the same variety. The tested varieties were subsequently categorized into four levels of resistance (Grades 1–4). The variety GT31 and LC05-136 showed the highest resistance (Grade 1), with a pest index of less than 0.35, while 12 varieties including FN91-21 and FN39 had a pest index of 0.35–0.45 (Grade 2). Five varieties including YT60 and YZ03-194 had a pest index of 0.45–0.55 (Grade 3), while FN38 and DZ03-83 had the lowest resistance (Grade 4), with a pest index of greater than 0.65. Correlation analysis further revealed that the number of thrips present on a single plant was related to the level of resistance; however, the correlation was relatively weak. These findings provide a preliminary evaluation of thrips resistance screening in sugarcane, laying the foundation for analysis of thrips-resistant genes, breeding of thrips-resistant varieties and resistance evaluation in new sugarcane germplasms.

Sugarcane streak mosaic virus (SCSMV) is one of the most important pathogens associated with sugarcane mosaic disease. In recent years, the rapid spread of SCSMV in Yunnan province, China, has raised concerns. In this study, we analyzed 110 sugarcane samples with mosaic symptoms taken from main growing areas in Yunnan province. RT-PCR detection results showed that 94% samples were positive for SCSMV. We subsequently analyzed the genetic diversity and population structure of SCSMV based on the coat protein (CP) sequences of 48 isolates. The overall mean value of nucleotide diversity for all isolates was 0.01127 ± 0.00082. Phylogenetic analysis based on CP gene sequences showed that all the Chinese isolates were clustered into one group, showing a distinct geographic feature. Analysis of population structure showed that the population from this study is not genetically different from that in China, but is genetically different from populations from other countries. Our results suggest that SCSMV has become the main pathogen of sugarcane mosaic disease in Yunnan province, China and that the recent rapid spread of SCSMV in Yunnan is not due to the invasion of new SCSMV populations.

Sugarcane mosaic disease is one of the most serious and prevalent viral diseases of sugarcane in China. The use of resistant varieties is the most economical and effective measures for controlling this disease. Sugarcane streak mosaic virus (SCSMV) and Sorghum mosaic virus (SrMV) are the two predominant pathogens of mosaic disease in the cane-growing regions of China. In 2015, 2016, double resistance to SCSMV and SrMV was identified once a year using a combination of artificial stalk-cutting inoculation and RT-PCR detection in 71 new elite sugarcane varieties/clones. Of the 71 new elite sugarcane varieties/clones, 24 were highly (Grade 1) to moderately resistant (Grade 3), and 47 were susceptible (Grade 4) to highly susceptible (Grade 5) to SCSMV. Furthermore, 27 were highly (Grade 1) to moderately resistant (Grade 3), and 44 were susceptible (Grade 4) to highly susceptible (Grade 5) to SrMV. Fifteen new elite sugarcane varieties/clones were highly resistant (Grade 1) to resistant (Grade 2) to SCSMV and SrMV. While, five new elite sugarcane varieties/clones (Yuegan 34, Yuetang 55, Yunzhe 03–258, Yunzhe 05–51, Yunzhe 06–80) were highly resistant (Grade 1) to SCSMV and SrMV. We defined the resistance of 71 new elite sugarcane varieties/clones to the two main pathogens of mosaic disease and identified 15 resistant to both SCSMV and SrMV. These results provide an elite resistance resource for effective prevention and control of mosaic disease and may serve as a reference for commercial varieties.

Background Insects cannot synthesize sterols and must acquire them from food. The mechanisms underlying how insects uptake dietary sterols are largely unknown except that NPC1b, an integral membrane protein, has been shown to be responsible for dietary cholesterol uptake in Drosophila melanogaster. However, whether NPC1b orthologs in other insect species, particularly the economically important pests, function similarly remains to be determined. Results In this study, we characterized the function of NPC1b in Helicoverpa armigera, a global pest that causes severe yield losses to many important crops. Limiting dietary cholesterol uptake to insects significantly inhibited food ingestion and weight gain. Compared to the wild-type H. armigera, the CRISPR/Cas9-edited NPC1b mutant larvae were incapable of getting adequate cholesterol and died in their early life stage. Gene expression profile and in situ hybridization analyses indicated that NPC1b was mainly expressed in the midgut where dietary cholesterol was absorbed. Expression of NPC1b was also correlated with the feeding life stages and was especially upregulated during early larval instars. Protein-ligand docking and sequence similarity analyses further demonstrated that NPC1b proteins of lepidopteran insects shared a relatively conserved cholesterol binding region, NPC1b_NTD, which, however, was highly divergent from bees-derived sequences. Conclusion NPC1b was crucial for dietary cholesterol uptake and growth of H. armigera, and therefore could serve as an insecticide target for the development of a novel pest-management approach to control this economically significant insect pest with little off-target effect on bees and sterol-autotrophic animals. © 2020 Society of Chemical Industry.

Brown rust, caused by Puccinia melanocephala has become a serious problem for sugarcane production in China. Therefore, 50 new elite varieties bred by the Chinese Sugarcane System were screened for brown rust resistance and the presence of a major resistance gene, Bru 1. Identification of field resistance and molecular detection of the brown rust resistance gene Bru 1 in the 50 new elite varieties and 2 main cultivated varieties were carried out in 2014 and 2015 at regional experimental stations of the China Sugarcane System in Dehong and Baoshan, Yunnan Province, where the incidence of brown rust is particularly high. Resistance evaluations based on ratings of symptom severity resulting from field infection showed that 32 (64%) of the 50 new elite varieties were highly resistant (n = 13, 26%), resistant (n = 16, 32%) and moderately resistant (n = 3, 6%). Molecular detection identified Bru 1 in 27 (54%) of these resistant varieties suggesting that brown rust resistance in these new elite varieties is primarily controlled by Bru 1. The absence of Bru 1 in the remaining five resistant suggested the presence of additional brown rust resistance genes. Two main cultivated varieties were highly resistant and Bru 1 - positive varieties. Different series differed in the frequencies of resistant and Bru 1 - positive varieties. Yunzhe series varieties had the highest frequency of resistant varieties (87.5%) and the highest frequency of Bru 1-positive varieties (81.3%); In contrast, the Yuetang series varieties had the lowest frequency of resistant varieties (40%) and the lowest frequency of Bru 1-positive varieties (30%). In this study, we defined the field resistance of 50 new elite varieties to brown rust, and determined the distribution of brown rust resistant gene Bru 1 in different series varieties. These results will provide new options for commercial production with effective resistance to brown rust. Moreover, these new elite resistant varieties can be used for subsequent breeding and selection to produce agronomically superior varieties with effective brown rust resistance.

To confirm the phytoplasma association in 13 sugarcane white leaf (SCWL) symptomatic samples collected from Puer, China, nested PCR assays with phytoplasma universal primer pairs was performed. A ~ 1240 bp product was obtained from 10 out of 13 symptomatic samples, and 16S rRNA gene sequencing results showed that all sequences of SCWL isolates were 100% identical to each other. BLASTn analysis revealed 100% sequence similarity with the 16S rRNA genes from SCWL phytoplasma strains LC7 and LC9 from Lincang, Yunnan province, China belonging to 16SrXI-B subgroup. Virtual restriction fragment length polymorphism also confirmed that Puer SCWL strains were member of the 16SrXI-B subgroup.

In 2015, symptoms of red stripe were first viewed on leaves of sugarcane plants growing in Yuanjiang, Yunnan Province. Symptomatic leaf samples were collected from infected plants and isolations were made on NA medium. Pure culture isolates from resulting single colony were identified based on cultural and morphological characteristics as Acidovorax avenae subsp. avenae. Morphological identification was confirmed by sequence analysis of 16S rDNA. Pathogenicity tests confirmed Yunnan isolates of Acidovorax avenae subsp. avenae as the causal agent of red stripe on sugarcane. This is the first report of this disease in Yuanjiang, Yunnan, China. Field surveys showed that the sugarcane cultivar ‘Yunzhe03-194’ was highly susceptible, and disease incidence varied from 8 to 80%. Nine other sugarcane cultivars were found to be symptomless in the field. Further research is required to confirm the resistance levels of the sugarcane varieties.

Seventy-seven mosaic leaf samples from new sugarcane varieties from national regional trials in Kaiyuan, Mile and Yuanjiang, Yunnan Province, China, were collected in 2015 and analyzed using reverse-transcription polymerase chain reaction (RT-PCR) to determine the pathogen causing mosaic symptoms in sugarcane. Three viruses, Sugarcane streak mosaic virus (SCSMV), Sorghum mosaic virus (SrMV) and Sugarcane mosaic virus (SCMV), were detected in 100, 27.3 and 1.3% of the samples respectively. Co-infection with SCSMV, SrMV and SCMV was also found for the first time in the new variety Qiantang 5. Nucleotide sequences of the coat protein (CP) gene of 48 SCSMV and 17 SrMV isolates were subsequently sequenced and analyzed with the CP gene sequences of SCSMV and SrMV reported in GenBank to perform phylogenetic analyses. Geographic differences were observed in the phylogenetic tree of SCSMV and all SCSMV isolates from China clustered into a single China group. Moreover, SCSMV isolates from different new varieties were distributed across all subgroups or clustered into an independent branch in the China group. SrMV isolates obtained in this study and those published in GenBank clustered into two groups and were distributed into different subgroups within these groups. Furthermore, the distribution of SrMV isolates from different new varieties overlapped in these different groups and subgroups, with no obvious geographic differences. These results provide a scientific basis for sugarcane mosaic or streak mosaic disease resistance breeding and effective control of this disease.

Brown rust caused by Puccinia melanocephala H. Sydow & P. Sydo is an important sugarcane disease that severely threatens sugarcane production in China. Breeding and planting resistant cultivars is the most cost-effective and efficient strategy for the control the disease. In addition, exploration, research, and utilization of resistant genes are the basis for resistance breeding. In the present study, four sugarcane varieties without the Bru1 gene and highly resistant to brown rust were used as male parents, and four varieties highly susceptible to brown rust were used as the female parents to configure hybrids. Six F1 hybrid populations were subjected to authenticity determination using SSR markers, brown rust resistance phenotype identification using artificial inoculation, and molecular detection of Bru1 gene. The results showed that the genetic segregation ratios in F1 hybrid populations of Yuetang 03-393 (HS) × ROC24 (HR) and Liucheng 03-1137 (HS) × Dezhe 93-88 (HR) were 3R:1S and 1R:3S, respectively, and Bru1 gene was not detected in their F1 hybrids. The results indicated that two dominant novel genes controlled brown rust resistance in Yuetang 03-393 (HS) × ROC24 (HR) and two recessive novel genes controlled brown rust resistance in Liucheng 03-1137 (HS) × Dezhe 93-88 (HR). Two genetically segregating populations that could be used for the localization of novel brown rust resistance genes were developed in the present study, and could lay a suitable foundation and provide theoretical reference for the analysis of the genetic mechanism of brown rust resistance, genetic map construction, mapping of novel resistance genes and the development of closely associated molecular markers in the future.

To improve the thermostability of the lipase LIP2 from Yarrowia lipolytica, molecular dynamics (MD) simulations at various temperatures were used to investigate the common fluctuation sites of the protein, which are considered to be thermally weak points. Two of these residues were selected for mutations to improve the enzyme's thermostability, and the variants predicted by MD simulations to have improved thermostability were expressed in Pichia pastoris GS115 for further investigations. According to the proline rule, the high fluctuation site S115 or V213 was replaced with proline residue, the two lipase mutants S115P and V213P were obtained. The mutant V213P exhibited evidently enhanced thermostability with an approximately 70% longer half-life at 50 °C than that of the parent LIP2 expressed in P. pastoris. The temperature optimum of V213P was 42 °C, which was about 5.0 °C higher than that of the parent LIP2, while its specific catalytic activity was comparable to that of the parent and reached 876.5 U/mg. The improved thermostability of V213P together with its high catalytic efficiency indicated that the rational design strategy employed here can be efficiently applied for structure optimization of industrially important enzymes.

Sugarcane brown rust induced by Puccinia melanocephala is an important global disease. Bru1 is a major brown rust resistance gene and has been shown to provide broad-spectrum resistance against various brown rust pathogen strains originating from different countries. Thus, brown rust resistance genes have important implications for sugarcane brown rust resistance breeding programs. The objectives of this study were to (1) assess the level of resistance of common sugarcane crossbreeding parents to P. melanocephala, (2) determine the distribution of the Bru1 gene in common sugarcane crossbreeding parents in China, (3) screen, evaluate, and select rust-resistant parental lines to improve the efficiency of breeding for brown rust resistance. Brown rust resistance in 101 common sugarcane breeding parents was collected from the Chinese National Nursery of Sugarcane Germplasm Resources and was scored and identified at the seedling stage and was assayed for the presence of the brown rust resistance gene Bru1. The results showed that Bru1 was detected in 48 (47.5 %) resistant clones out of the 101 tested parental lines, indicating that brown rust resistance in common sugarcane breeding parents in China is primarily controlled by Bru1. The apparent absence of Bru1 from the other 29 resistant and 24 susceptible parental lines suggested that they may carry brown rust resistance-associated genes other than Bru1. These results may facilitate future efforts to breed varieties with improved resistance to brown rust and may provide additional resistant parental lines for selecting and planting elite-resistant varieties to effectively control brown rust disease in sugarcane.

Sugar cane white leaf (SCWL) is a serious disease caused by phytoplasmas. In this study, we performed nested PCR with phytoplasma universal primer pairs (P1/P7 and R16F2n/R16R2) for the 16S rRNA gene to detect SCWL phytoplasmas in 31 SCWL samples collected from Baoshan and Lincang, Yunnan, China. We cloned and sequenced the nested PCR products, revealing that the 16S rRNA gene sequences from 31 SCWL samples were all 1247 bp in length and shared more than 99 % nucleotide sequence similarity with the 16S rRNA gene sequences of SCWL phytoplasmas from various countries. Based on the reported 16S rRNA gene sequence data from SCWL isolates of various countries, we conducted phylogenetic and virtual RFLP analysis. In the resulting phylogenetic tree, all SCWL isolates clustered into two branches, with the Lincang and Baoshan SCWL phytoplasma isolates belonging to different branches. The virtual RFLP patterns show that phytoplasmas of the Lincang branch belong to subgroup 16SrXI-B. However, the virtual RFLP patterns revealed by HaeIII digestion of phytoplasmas of the Baoshan branch differed from those of subgroup 16SrXI-B. According to the results of phylogenetic and virtual RFLP analysis, we propose that the phytoplasmas of the Baoshan branch represent a new subgroup, 16SrXI-D. These findings suggest that SCWL is caused by phytoplasmas from group 16SrXI, including subgroup 16SrXI-B and a new subgroup, 16SrXI-D.

Acute infections with enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) usually cause Hand, foot and mouth disease (HFMD) among infants and young children with several large outbreaks worldwide. Unfortunately, the molecular mechanisms underlying enterovirus infections remain largely unknown. In this study, we analyzed the genome-wide DNA methylation patterns of host cells in response to EV71 and CVA16 infections using the Illumina Infinium HumanMethylation450 BeadChip. Of over 480,000 loci studied, significant differential methylation was observed between EV71 infected-cells and control cells at 3957 CpG sites, out of which 2478 were hypermethylated and 1479 were hypomethylated, whereas CVA16 infection resulted in methylation level changes of 5194 CpG sites with 4288 hypermethylated and 906 hypomethylated. These differential methylated loci displayed a wide range of genomic distributions in chromosomes, inside and surrounding areas (shores and shelves) of CpG islands, as well as functional gene regions including promoter, gene body and 3'UTR. Based on methylation alterations, 1189 genes were identified to be potentially co-associated with the replication processes of two enteroviruses. GO function annotation and enrichment analysis of 1189 common differentially methylated genes reflected a broad spectrum of biological regulatory events during viral infection. KEGG pathway analysis indicated the involvement of diverse signaling pathways including viral myocarditis, Notch signaling and antigen processing and presentation. Our present study provides a novel insight into enterovirus-host interaction network at epigenetic profile, thus contributing to improved understanding of HFMD pathogenesis.

For many years, the control and treatment of sugarcane pests in China mainly relies on highly toxic and broad-spectrum chemical pesticides, such as methamidophos and omethoate. As a result, many nontargets, natural enemy species rapid declines, the dynamic relationship between pests and their natural enemies were disrupted, and pests develop resistance, natural enemy population decreased. It alleviates the function on natural factor of controlling pests and makes the prevention and control work more passive. Moreover, the misuse of pesticides leads to pesticide residue, environment pollution, and human health harmfulness. Therefore, a new task for plant protection workers is to control the pests in a comprehensive and environmentally friendly way. In recent years, biological control has become one of the important measures in integrated pest control, and its social and ecological benefits are attracting more and more attention from the state and society (Pu 1984; Bao 1988; Pu and Li 1996; Zhou et al. 1999). The study and development of using dominant natural enemies to control the pests can get the obvious effect on increasing yield, reducing environmental pollution, maintaining ecological balance, saving energy, and cutting down production cost (Yang et al. 2006; An et al. 2009). With China’s accession to the WTO, it is becoming increasingly important to protect and utilize the natural enemies of sugarcane pests, when the implementation of the process of quality standardization of agricultural products has been continuously promoted.

There are more than 120 sugarcane diseases that have been found in the world presently (Chen 1982; Rott et al. 2000), and more than 60 have been reported in China (The important sugarcane diseases research cooperation group 1991; Lu et al. 1997; Huang and Li 2014, 2016). Mastering the disease types, damage, and distribution in the sugarcane area can provide scientific basis for the sugarcane disease-resistance breeding, quarantine of introduced varieties, and disease management and research. Sugarcane production areas in China have drought in winter and spring and hot climate and good rainfall in summer and autumn. With the rise of temperature in spring, in case of cold current as well as the rainfall and temperature decline and the poor management, planted sugarcane in winter and this season are easily infected by pineapple disease and the ratoon crops are easily damaged by red rot disease. Because the whole seedling stage is in the dry season, smut becomes substantial causing sparse breakup. After June, with the high temperature and humidity, eyespot, brown stripe, yellow spot, pokkah boeng, rust, ring spot, and leaf scorch diseases are easily occurring and epidemic. Especially in recent years, introducing seedcanes frequently and transposing seedcanes in sugarcane planting area make some dangerous seedcane-transmitted diseases (ratoon stunting disease, mosaic, smut, yellow leaf syndrome, etc.) which are spread by seedcane between sugarcane areas. Then, caused sugarcane diseases occur commonly and damage more severely; have a serious impact on sugarcane yield, quality, and ratooning capacity; and bring serious hidden trouble to the safe production of sugarcane. In order to scientifically control the spread and damage of sugarcane disease, enhancing the capabilities of disaster prevention and reduction and ensuring the quality of sugarcane variety and safety of sugarcane production in 20 diseases that commonly occur in sugarcane production are systematically described in this chapter with clear-colored photos and scientific, accurate words. The content includes the occurrence and damage, identification of symptoms, characteristics of epidemic infection, and control measures.

This study investigated the composition and proportions of fecal microbiota in Sprague-Dawley rats after consuming two genetically modified (GM) corn lines in comparison with the isogenic corn and the AIN93G standard feed for 10 weeks using bar-coded 16S rRNA gene sequencing. As a result, GM corn did not significantly alter the overall health and alpha-diversity of fecal microbiota. Fecal microbiota structures could be separated into noncorn and corn but not non-GM and GM corn subgroups. Both non-GM and GM corn caused the increase in bacterial populations related to carbohydrates utilization, such as Lactobacillus, Barnesiella, and Bifidobacterium, and the reduction in potentially pathogenic populations, such as Tannerella and Moraxellaceae. In conclusion, similar effects on the fecal microbiota were observed after consuming a GM- and non-GM-corn-based diet for long periods. Further studies are warranted to elucidate the functional relevance of the changes in the proportions of bacterial populations in these diets.

Ficellomycin is an aziridine antibiotic produced by Streptomyces ficellus, which displays high in vitro activity against Gram-positive bacteria including multidrug resistant strains of Staphylococcus aureus. Compared to currently available antibiotics, ficellomycin exhibits a unique mechanism of action—it impairs the semiconservative DNA replication by inducing the formation of deficient 34S DNA fragments, which lack the ability to integrate into larger DNA pieces and eventually the complete bacterial chromosome. Until recently, some important progress has been made in research on ficellomycin synthesis and biosynthesis, opening the perspective to develop a new generation of antibiotics with better clinical performance than the currently used ones. In this review, we will cover the discovery and biological activity of ficellomycin, its biosynthesis, mode of action, and related synthetic analogs. The role of ficellomycin and its analogs as an important source of drug prototypes will be discussed together with future research prospects.

Sugarcane white leaf (SCWL) phytoplasma was identified from sugarcane varieties in fields of Lincang and Baoshan, and imported sugarcane varieties from Myanmar, Philippines and Thailand in quarantine glasshouse at Yunnan Province, China. The associated phytoplasmas were characterized as strains of 16SrXI group by using MLOX/MLOY and P1/P2 primer pairs, 16Sr sequence identity and phylogenetic relationships.

Biohydrogen production from the pulp and paper effluent containing rich lignocellulosic material could be achieved by the fermentation process. Xylose, an important hemicellulose hydrolysis product, is used less efficiently as a substrate for biohydrogen production. Moreover, azo dyes are usually added to fabricate anticounterfeiting paper, which further increases the complexity of wastewater. This study reports that xylose could serve as the sole carbon source for a pure culture of Klebsiella oxytoca GS-4-08 to achieve simultaneous decolorization and biohydrogen production. With 2 g liter −1 of xylose as the substrate, a maximum xylose utilization rate (UR xyl ) and a hydrogen molar yield (HMY) of 93.99% and 0.259 mol of H 2 mol of xylose −1 , respectively, were obtained. Biohydrogen kinetics and electron equivalent ( e − equiv) balance calculations indicated that methyl red (MR) penetrates and intracellularly inhibits both the pentose phosphate pathway and pyruvate fermentation pathway, while methyl orange (MO) acted independently of the glycolysis and biohydrogen pathway. The data demonstrate that biohydrogen pathways in the presence of azo dyes with sulfonate and carboxyl groups were different, but the azo dyes could be completely reduced during the biohydrogen production period in the presence of MO or MR. The feasibility of hydrogen production from industrial pulp and paper effluent by the strain if the xylose is sufficient was also proved and was not affected by toxic substances which usually exist in such wastewater, except for chlorophenol. This study offers a promising energy-recycling strategy for treating pulp and paper wastewaters, especially for those containing azo dyes. IMPORTANCE The pulp and paper industry is a major industry in many developing countries, and the global market of pulp and paper wastewater treatment is expected to increase by 60% between 2012 and 2020. Such wastewater contains large amounts of refractory contaminants, such as lignin, whose reclamation is considered economically crucial and environmentally friendly. Furthermore, azo dyes are usually added in order to fabricate anticounterfeiting paper, which further increases the complexity of the pulp and paper wastewater. This work may offer a better understanding of biohydrogen production from xylose in the presence of azo dyes and provide a promising energy-recycling method for treating pulp and paper wastewater, especially for those containing azo dyes.

Sugarcane white leaf (SCWL) is an important disease caused by a phytoplasma. In Baoshan, Yunnan, China, SCWL was firstly observed in 2012, and has extended its area of occurrence to 600 hm² . Up to 52% of the plants may become diseased in a field and even complete loss of cane yield may result in the heavily infected fields, posing a serious threat to Yunnan sugar industry. To ascertain the causal agent of suspected SCWL disease in Yunnan, nested PCR using two sets of phytoplasma primer pairs (MOLX/MLOY and P1/ P2) was used to successfully amplify a genomic region of the 16S ribosomal DNA (16S rDNA) from 36 suspected SCWL samples. On the basis of sequencing, phylogenetic analysis and nucleotide alignments of 17 nested PCR products randomly selected from positive samples, identical fragments of 210bp in length were obtained that could be clustered into the 6Sr group XI (Rice Yellow Dwarf group) and shared 100% identity with the 16S-23S intergenic spacer region (ISR) of a member of this group, the SCWL phytoplasma (GenBank: HQ917068), and 99.52% with Sugarcane grassy shoot phytoplasma, of the same group. These results indicate that the SCWL disease in Baoshan is caused by a phytoplasma of the 6Sr group XI.

Exploring and utilizing resistant germplasm resources plays a pivotal role in breeding for disease resistance, while screening resistant germplasm is important for selecting and breeding varieties resistant to disease. In the present study, we used PCR to detect the ratoon stunting disease (RSD) bacterium Leifsonia xyli subsp. xyli (Lxx) in 137 sugarcane core germplasms from the National Nursery of Sugarcane Germplasm Resources (NNSGR, Kaiyuan, China) in 2009, 2010 and 2011. A total of 21 germplasms that tested negative for Lxx in 2009 and 2010 were selected for further Lxx detection after being subjected to artificial inoculation in 2011 and 2012. The 21 core germplasms that were negative for Lxx after natural infection and artificial inoculation can provide elite resistance source materials and reference frames for the effective breeding of RSD-resistant sugarcane varieties. Under natural conditions, 116 (84.67%) and 21 (15.33%) out of 137 germplasms were positive and negative for Lxx, respectively, as determined by PCR detection, which suggests that a relatively high ratio of sugarcane core germplasms was sensitive to RSD, while few were resistant to RSD. The sequencing and analysis of 30 randomly selected PCR products showed that all 30 sequences were identical or highly homologous to the corresponding Lxx genome region published in GenBank (99.54–100% similarity). Lxx can be detected effectively and precisely by PCR. We therefore recommend PCR as a rapid, low cost and simple procedure to score sugarcane core germplasms for RSD resistance.

The introduction of the sugarcane white leaf (SCWL) phytoplasma into Yunnan Province, China was confirmed for the first time by nested PCR. Sequencing and analysis of 17 nested PCR products of the 16S rDNA of the phytoplasma from random samples showed that these sequences were identical. A BLAST search indicated that the sequences were highly homologous (99.05–100% similarity) to the corresponding genome region in GenBank. Field surveys showed that disease incidence among affected cultivars was 13–52% and up to 100% in seriously infected fields. The sugarcane cultivar ‘Yuetang86-368’ was highly susceptible. SCWL is a destructive disease on sugarcane that requires preventive quarantine management measures to prevent its further spread.

Pacific white shrimp Litopenaeus vannamei (1050 individuals with initial weight of 1.01 ± 0.001 g) were fed either control diet or one of six dietary astaxanthin (AX) concentration (25, 50, 75, 100, 125 and 150 mg kg−1) diets for 56 days in 35 tanks (30 shrimp per tank). After 56 days of culture, shrimp-fed AX125 and AX150 diets had higher (P

In order to investigate the effects of lysine and dissolved oxygen on grass carp, the grass carp were fed with 13, 15 and 17 g kg −1 lysine diet at about 6 mg L−1 (high dissolved oxygen, HO group) or 3.5 mg L−1 (low dissolved oxygen, LO group) dissolved oxygen level, for 8 weeks. The fish were fed to apparent satiation by hand. The results showed that apparent digestibility of protein, energy and dry matter were decreased significantly when grass carp were fed at 3.5 mg L−1 dissolved oxygen, and feed intake (FI) was also inhibited by low dissolved oxygen (P

In 2010 and 2011, 70 wild sugarcane germplasms including 37 Saccharum spontaneum and 33 Erianthus arundinaceus from the National Nursery of Sugarcane Germplasm Resources (NNSGR, Kaiyuan, China) were screened for their resistance to sugarcane mosaic disease caused by Sorghum mosaic virus (SrMV-HH, GenBank accession number DQ530434) by dropping inoculum on stem-section and SrMV RT-PCR detection. The results showed that of the 70 tested germplasms, 39 (55.7%) were highly (Grade 1) to moderately resistant (Grade 3) to SrMV. E. arundinaceus had more resistant lines than S. spontaneum. Of the 33 tested E. arundinaceus germplasms, 22 (66.7%) were highly (Grade 1) to moderately resistant (Grade 3), while 17 (45.9%) of the 37 tested S. spontaneum germplasms were highly (Grade 1) to moderately resistant (Grade 3) to SrMV. The results suggested that S. spontaneum and E. arundinaceus from the NNSGR contain valuable germplasms resistant to SrMV, and the screened germplasms in the present study could provide promising resistance sources for sugarcane breeding.

This experiment was conducted to investigate the effect of ghrelin and des-ghrelin on feed intake, growth performance and hypothalamic neuropeptide Y (NPY) mRNA expression of grouper, Epinephelus coioides . Ghrelin and des-ghrelin (non-acylated form) were supplemented in the basal diet at the dose of 0, 2, 4 mg kg − 1 . The highest feed intake, weight gain (WG, %) and final body weight (FBW, g) were recorded when ghrelin was added at the dose of 4 mg kg − 1 and they were significantly different from that of the control ( P P > 0.05) effect on growth performance and feed intake was observed in fish fed diet supplemented with des-ghrelin at 2 and 4 mg kg − 1 . There were no significant differences ( P > 0.05) in the proximate composition (crude protein, crude lipid, ash and moisture), slaughter variables (condition factor, viscerasomatic index, hepatosomatic index and intraperitoneal fat ratio) and serum lipid (total cholesterol, triglyceride, low density lipoprotein and high density lipoprotein) in fish fed all the test diets. Grouper hypothalamic NPY mRNA expression was significantly ( P − 1 ) was supplemented in the diet. However, grouper fed the diet added with des-ghrelin (2, 4 mg kg − 1 ) did not show an increase ( P > 0.05) in the expression of hypothalamic NPY mRNA. The present results indicated for the first time that ghrelin supplemented in the diet could stimulate feed intake, growth and hypothalamic NPY mRNA expression of grouper, while des-ghrelin did not have those effects.

A 63-day growth trial was undertaken to estimate the effects of supplemented lysine and methionine with different dietary protein levels on growth performance and feed utilization in Grass Carp (Ctenopharyngodon idella). Six plant-based practical diets were prepared, and 32CP, 30CP and 28CP diets were formulated to contain 320 g kg−1, 300 g kg−1 and 280 g kg−1 crude protein without lysine and methionine supplementation. In the supplementary group, lysine and methionine were added to formulate 32AA, 30AA and 28AA diets with 320 g kg−1, 300 g kg−1 and 280 g kg−1 dietary crude protein, respectively, according to the whole body amino acid composition of Grass Carp. In the groups without lysine and methionine supplementation, weight gain (WG, %) and specific growth rate (SGR, % day−1) of the fish fed 32CP diet were significantly higher than that of fish fed 30CP and 28CP diets, but no significant differences were found between 30CP- and 28CP-diet treatments. WG and SGR of the fish fed 32AA and 30AA diets were significantly higher than that of fish fed 28AA diets, and the performance of grass carp was also significantly improved when fed diets with lysine and methionine supplementation (P