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Journal of Integrative Agriculture
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Development of a reverse-transcription loop-mediated isothermal amplification assay to detect avian influenza viruses in clinical specimens
SHI Lin, YU Xue-wu, YAO Wei, YU Ben-liang, HE Li-kun, GAO Yuan, ZHANG Yun-xian, TIAN Guo-bin, PING Ji-hui, WANG Xiu-rong
2019, 18 (
7
): 1428-1435. DOI:
10.1016/S2095-3119(19)62700-0
Abstract
(
249
)
PDF in ScienceDirect
In recent years, the avian influenza has brought not only serious economic loss to the poultry industry in China but also a serious threat to human health because of the avian influenza virus (AIV) gene recombination and reassortment. Until now, traditional RT-PCR, fluorescence RT-PCR and virus isolation identification have been developed and utilized to detect AIV, but these methods require high-level instruments and experimental conditions, not suitable for the rapid detection in field and farms. In order to develop a rapid, sensitive and practical method to detect and identify AIV subtypes, 4 specific primers to the conserved region of AIV M gene were designed and a loop-mediated isothermal amplification (RT-LAMP) method was established. Using this method, the M gene of H1–H16 subtypes of AIV were amplified in 30 min with a water bath and all 16 H subtypes of AIV were able to be visually identified in presence of fluorescein, without cross reaction with other susceptible avian viruses. In addition, the detection limit of the common H1, H5, H7, and H9 AIV subtypes with the RT-LAMP method was 0.1 PFU (plaque-forming unit), which was 10 times more sensitive than that using the routine RT-PCR. Further comparative tests found that the positivity rate of RT-LAMP on detecting clinical samples was 4.18% (14/335) comparing with 3.58% (12/335) from real-time RT-PCR. All these results suggested that the RT-LAMP method can specifically detect and identify AIV with high sensitivity and can be considered as a fast, convenient and practical method for the clinic test and epidemiological investigation of AIV.
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Molecular mapping of leaf rust resistance genes in the wheat line Yu 356-9
HAN Liu-sha, LI Zai-feng, WANG Jia-zhen, SHI Ling-zhi, ZHU Lin, LI Xing, LIU Da-qun, Syed J A Shah
2015, 14 (
7
): 1223-1228. DOI:
10.1016/S2095-3119(14)60964-3
Abstract
(
2360
)
PDF in ScienceDirect
The Chinese wheat line Yu 356-9 exhibits a high level of resistance to leaf rust. In order to decipher the genetic base of resistance in Yu 356-9, gene postulation, inheritance analyses, and chromosome linkage mapping were carried out. Gene postulation completed using 15 leaf rust pathotypes and 36 isogenic lines indicated that Yu 356-9 was resistant to all pathotypes tested. F1 and F2 plants from the cross Yu 356-9 (resistant)/Zhengzhou 5389 (susceptible) were tested with leaf rust pathotype “FHNQ” in the greenhouse. Results indicated a 3:1 segregation ratio, indicative of the presence of a single dominant leaf rust resistance gene in Yu 356-9 which was temporarily designated as LrYu. Bulk segregant analysis and molecular marker assays were used to map LrYu. Five simple sequence repeat (SSR) markers on chromosome 2BS were found closely linked to LrYu. Among these markers, Xwmc770 is the most closely linked, with a genetic distance of 5.7 cM.
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Soil phosphorus dynamic, balance and critical P values in longterm fertilization experiment in Taihu Lake region, China
SHI Lin-lin, SHEN Ming-xing, LU Chang-yin, WANG Hai-hou, ZHOU Xin-wei, JIN Mei-juan, WU Tong-dong
2015, 14 (
12
): 2446-2455. DOI:
10.1016/S2095-3119(15)61183-2
Abstract
(
1796
)
PDF in ScienceDirect
Phosphorus (P) is an important macronutrient for plant but can also cause potential environmental risk. In this paper, we studied the long-term fertilizer experiment (started 1980) to assess the soil P dynamic, balance, critical P value and the crop yield response in Taihu Lake region, China. To avoid the effect of nitrogen (N) and potassium (K), only the following treatments were chosen for subsequent discussion, including: C0 (control treatment without any fertilizer or organic manure), CNK treatment (mineral N and K only), CNPK (balanced fertilization with mineral N, P and K), MNK (integrated organic manure and mineral N and K), and MNPK (organic manure plus balanced fertilization). The results revealed that the response of wheat yield was more sensitive than rice, and no significant differences of crop yield had been detected among MNK, CNPK and MNPK until 2013. Dynamic and balance of soil total P (TP) and Olsen-P showed soil TP pool was enlarged significantly over consistent fertilization. However, the diminishing marginal utility of soil Olsen-P was also found, indicating that high-level P application in the present condition could not increase soil Olsen-P contents anymore. Linear-linear and Mitscherlich models were used to estimate the critical value of Olsen-P for crops. The average critical P value for rice and wheat was 3.40 and 4.08 mg kg–1, respectively. The smaller critical P value than in uplands indicated a stronger ability of P supply for crops in this paddy soil. We concluded that no more mineral P should be applied in rice-wheat system in Taihu Lake region if soil Olsen-P is higher than the critical P value. The agricultural technique and management referring to activate the plant-available P pool are also considerable, such as integrated use of low-P organic manure with mineral N and K.
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