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SLAF marker based QTL mapping of fruit-related traits reveals a major-effect candidate locus ff2.1 for flesh firmness in melon
CHEN Ke-xin, DAI Dong-yang, WANG Ling, YANG Li-min, LI Dan-dan, WANG Chao, JI Peng, SHENG Yun-yan
2023, 22 (11): 3331-3345.   DOI: 10.1016/j.jia.2023.02.014
Abstract166)      PDF in ScienceDirect      

Flesh firmness (FF) is an important and complex trait for melon breeders and consumers.  However, the genetic mechanism underlying FF is unclear.  Here, a soft fruit melon (P5) and a hard fruit melon (P10) were crossed to generate F2, and the FF and fruit-related traits were recorded for two years.  By performing quantitative trait locus (QTL) specific-locus amplified fragment (SLAF) (QTL-SLAF) sequencing and molecular marker-linkage analysis, 112 844 SLAF markers were identified, and 5 919 SNPs were used to construct a genetic linkage map with a total genetic distance of 1 356.49 cM.  Ten FF- and fruit-related QTLs were identified.  Consistent QTLs were detected for fruit length (FL) and fruit diameter (FD) in both years, and QTLs for single fruit weight (SFW) were detected on two separate chromosomes in both years.  For FF, the consistent major locus (ff2.1) was located in a 0.17-Mb candidate region on chromosome 2.  Using 429 F2 individuals derived from a cross between P5 and P10, we refined the ff2.1 locus to a 28.3-kb region harboring three functional genes.  These results provide not only a new candidate QTL for melon FF breeding but also a theoretical foundation for research on the mechanism underlying melon gene function.

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Non-target-site and target-site resistance to AHAS inhibitors in American sloughgrass (Beckmannia syzigachne)
WANG Jing-jing, LI Xiang-ju, LI Dan, HAN Yu-jiao, LI Zheng, YU Hui-lin, CUI Hai-lan
2018, 17 (12): 2714-2723.   DOI: 10.1016/S2095-3119(18)62021-0
Abstract285)      PDF in ScienceDirect      
American sloughgrass (Beckmannia syzigachne (Steud.) Fernald) is one of the most competitive and malignant weeds in rice-wheat rotation fields in China.  American sloughgrass populations in the Jiangsu Province of China became less sensitive to acetohydroxyacid synthase (AHAS) inhibitors after repeated application for many years in these areas.  Two suspected resistant American sloughgrass populations (R1 and R2) collected in the field were detected the resistance to inhibitors of AHAS in whole-plant dose-response assays, compared to the susceptible (S) population.  These assays indicated that R1 showed low resistance to mesosulfuron-methyl (3.32-fold), imazapic (2.84-fold) and pyroxsulam (1.55-fold), moderate resistance to flazasulfuron (4.67-fold) and pyribenzoxim (7.41-fold), and high resistance to flucarbazone (11.73-fold).  However, using a combination of the cytochrome P450 inhibitor, malathion, with mesosulfuron-methyl resulted in a reduction in R1 resistance relative to mesosulfuron-methyl alone.  Furthermore, R2 was highly resistant to flazasulfuron (34.90-fold), imazapic (11.30-fold), flucarbazone (49.20-fold), pyribenzoxim (12.94-fold), moderately resistant to mesosulfuron-methyl (9.77-fold) and pyroxsulam (6.26-fold), and malathion had no effect on R2 resistance to mesosulfuron-methyl.  The full-length of AHAS genes was sequenced and the AHAS enzymes were assayed in vitro in order to clarify the mechanism of resistance to AHAS inhibitors in R1 and R2 populations.  The results demonstrated that R2 had a Pro-197-Ser mutation in the AHAS gene, and the sensitivity of R2 to the five AHAS inhibitors was decreased, which may result in R2 resistance to AHAS inhibitors.  There was no mutation in the AHAS gene of R1, and there were no significant differences in enzyme sensitivity between susceptible (S) and resistant (R1) populations.  An enhanced metabolism may be the main mechanism of R1 resistance to AHAS inhibitors.
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Effect of temperature on the development of sclerotia in Villosiclava virens
FAN Lin-lin, YONG Ming-li, LI Dan-yang, LIU Yi-jia, LAI Chao-hui, CHEN Hong-ming, CHENG Fang-min, HU Dong-wei
2016, 15 (11): 2550-2555.   DOI: 10.1016/S2095-3119(16)61400-4
Abstract3004)      PDF in ScienceDirect      
    The sclerotia of Villosiclava virens were found commonly in high altitude and the temperate regions, where the temperatures are relatively low in rice filling stage. To make sure if low temperature induce the sclerotial formation in V. virens, the inoculated rice panicles in laboratory and the diseased rice panicles cut from paddy fields were treated under different temperatures. The results showed that 3 days of night temperature at 15°C were enough to induce the sclerotial formation. The low temperature was much more effective for young balls with intact membranes. After appearance of chlamydospores on the ball surfaces, the sclerotium could not differentiate anymore. The sclerotia began to differentiate below the chlamydospore layer and gradually grew onto the ball surfaces. This suggests that low temperature in the early development stage of false smut balls is an important factor to induce the sclerotial differentiation, and rice cultivars with long growth periods are able to produce more sclerotium-bearing balls, which will produce mass of spores in paddy field in the coming year.
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Survey and examination of the potential alternative hosts of Villosiclava virens, the pathogen of rice false smut, in China
DENG Qi-de, YONG Ming-li, LI Dan-yang, LAI Chao-hui, CHEN Hong-ming, FAN Jing, HU Dong-wei
2015, 14 (7): 1332-1337.   DOI: 10.1016/S2095-3119(15)61030-9
Abstract2021)      PDF in ScienceDirect      
Rice false smut is caused by ascomycete Villosiclava virens, whose potential alternative hosts have been assumed previously. Here its potential alternative hosts were surveyed and identified from 2008 to 2013 in the main rice-growing regions in China. Two common weeds in paddy fields, Digitaria sanguinalis Scop. and Echinochloa crusgalli (L.) Beauv., were found to present the similar symptoms to smut diseases in a few individuals in 2012 and 2013 in Zhejiang and Sichuan provinces of China, respectively. After the examinations of the spore morphology, their infection and extension mode in hosts, pathogen cell wall components, and molecular identification, the two pathogens were identified to be the Basidiomycetes, Ustilago syntherismae and Ustilago trichophora, respectively. So far there has been no alternative host of V. virens to be identified in China. These suggest that the alternative hosts of V. virens, if they do exist, are not possible to play an important role in the pathogen life cycle and the disease epidemics.
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Rapid Recovery of Classical Swine Fever Virus Directly from Cloned cDNA
HUANG Jun-hua, LI Yong-feng, HE Fan, LI Dan, SUN Yuan, HAN Wen , QIU Hua-ji
2013, 12 (5): 877-883.   DOI: 10.1016/S2095-3119(13)60258-0
Abstract1675)      PDF in ScienceDirect      
The reverse genetics for classical swine fever virus (CSFV) is currently based on the transfection of in vitro transcribed RNA from a viral genomic cDNA clone, which is inefficient and time-consuming. This study was aimed to develop an improved method for rapid recovery of CSFV directly from cloned cDNA. Full-length genomic cDNA from the CSFV Shimen strain, which was flanked by a T7 promoter, the hepatitis delta virus ribozyme and T7 terminator sequences, was cloned into the lowcopy vector pOK12, producing pOKShimen-RzT . Direct transfection of pOKShimen-RzT into PK/T7 cells, a PK-15- derived cell line stably expressing bacteriophage T7 RNA polymerase, allowed CSFV to be rescued rapidly and efficiently, i.e., at least 12 h faster and 31.6-fold greater viral titer when compared with the in vitro transcription-based rescue system. Furthermore, the progeny virus rescued from PK/T7 cells was indistinguishable, both in vitro and in vivo, from its parent virus and the virus rescued from classical reverse genetics. The reverse genetics based on intracellular transcription is efficient, convenient and cost-effective. The PK/T7 cell line can be used to rescue CSFV directly from cloned cDNA and it can also be used as an intracellular transcription and expression system for studying the structure and function of viral genes.
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Conophylline Promotes the Proliferation of Immortalized Mesenchymal Stem Cells Derived from Fetal Porcine Pancreas (iPMSCs)
ZHANG Hui-ru, LI Dan, CAO Hui, Lü Xiao, CHU Yuan-kui, BAI Yao-fu, JIN Ya-ping, PENG
2013, 12 (4): 678-686.   DOI: 10.1016/S2095-3119(13)60286-5
Abstract1319)      PDF in ScienceDirect      
Conophylline, is a bis (indole) alkaloid consisting of two pentacyclic aspidosperma skeletons, isolated from Tabernaemontana divaricata, which has been found to induce b-cell differentiation in rat pancreatic acinar carcinoma cells and in cultured rat pancreatic tissue. However, the precise role of conophylline in the growth and survival of immortalized pancreatic mesenchymal stem cells (iPMSCs) derived from fetal porcine pancreas were not understood at present. To determine whether this molecule is involved in controlling the proliferation of iPMSCs, we examined the effects of conophylline on iPMSCs. We found that conophylline can robustly stimulate iPMSCs proliferation, even promote their potential differentiation into islet-like clusters analyzed by cell counting, morphology, RT-PCR and real-time PCR, Western blotting, glucose-stimulated insulin release and insulin content analysis. The effects of conophylline were inhibited by LY294002, which is the inhibitor of the PI3K pathway. These results suggest that conophylline plays a key role in the regulation of cell mass proliferation, maintenance of the undifferentiated state of iPMSCs and also promotes iPMSCs differentiated into insulin-producing cells.
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System fertilization improves soil quality and increases primary production in an integrated crop-livestock system
Vicente José Laamon Pinto Simões, Lóren Pacheco Duarte, Rafaela Dulcieli Daneluz Rintzel, Amanda Posselt Martins, Tales Tiecher, Leonardo Dallabrida Mori, Carolina Bremm, Marco Aurélio Carbone Carneiro, Paulo César de Faccio Carvalho
DOI: 10.1016/j.jia.2025.03.002 Online: 12 March 2025
Abstract4)      PDF in ScienceDirect      

Managing fertilization in integrated crop-livestock systems (ICLS) during periods of low nutrient export, known as system fertilization, can optimize nutrient use by enhancing the soil’s biochemical and physical-hydric properties. However, interdisciplinary studies on processes that improve input utilization in ICLS remain scarce. This study aimed to assess the relationships between the efficiencies of different nutrient management strategies in ICLS and pure crop systems (PCS) and the biochemical and physical-hydric quality of soil. Two fertilization strategies (system fertilization and crop fertilization) and two cropping systems (ICLS and PCS) were evaluated in a randomized block design with three replicates. In the PCS, soybean was grown followed by ryegrass as a cover crop. In the ICLS, sheep grazed on the ryegrass. In the crop fertilization, phosphorus and potassium were applied to the soybean planting, and nitrogen was applied in the ryegrass establishment. Nitrogen, phosphorus, and potassium were applied during ryegrass establishment in the system fertilization. Soil quality indexes were calculated using fourteen physical-hydric and biochemical soil indicators, and primary production and nutrient utilization efficiency were evaluated. System fertilization in ICLS enhanced the soil functions of water storage and availability for plants, structural stability, and resistance to degradation. System fertilization in ICLS improved the soil quality by 14% over PCS and 13% over crop fertilization in ICLS. Notably, this optimized system yielded the highest primary production. These findings underscore the pivotal role of system fertilization in ICLS to boost food production and enhance soil ecosystem services without increasing the consumption of external fertilizers. They advocate for a strategic shift towards system-level fertilization in integrated systems, and demonstrate for the first time in ICLS, the delicate balance between nutrient management, soil health, and sustainable productivity.

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