Multinucleate condition of Puccinia striiformis in colonies isolated from infected wheat leaves with macerating enzymes

1992 ◽  
Vol 70 (1) ◽  
pp. 222-224 ◽  
Author(s):  
J. Chong ◽  
Z. Kang ◽  
W. K. Kim ◽  
R. Rohringer

A fluorescence microscopy study was carried out to examine the multinuclear condition in parasitic colonies of one isolate of Puccinia striiformis in leaf tissue of a susceptible wheat. At 5 and 10 days after inoculation, colonies were isolated with macerating enzymes and sequentially stained with 4′-6-diamidino-2-phenylindole and Calcofluor for nuclear counts in the intercellular hyphal cells, haustorium mother cells, and urediniospores. In 10-day-old colonies, more than 98% of the hyphal cells located adjacent to the uredinia were binucleate. Elsewhere in the colonies, the hyphal cells were highly variable in nuclear number; approximately 20% contained two nuclei, 18% contained four, 26% contained six, and 28% contained eight. In 5-day-old colonies, the hyphal cells were also highly variable in nuclear number, regardless of where they were located in the colony. The nuclear condition of haustorium mother cells was less variable; the majority contained either two or four nuclei. All urediniospores examined were binucleate. In the parasitic state, the multinucleate condition apparently occurs only in P. striiformis and not in other cereal rust fungi. Key words: multinucleate condition, stripe rust, Puccinia striiformis, DAPI, Calcofluor.

2013 ◽  
Vol 103 (9) ◽  
pp. 927-934 ◽  
Author(s):  
Jie Zhao ◽  
Long Wang ◽  
Zhiyan Wang ◽  
Xianming Chen ◽  
Hongchang Zhang ◽  
...  

The wheat stripe rust pathogen (Puccinia striiformis f. sp. tritici) population in China has been reported to be a distinct genetic group with higher diversity than those in many other countries. Genetic recombination in the P. striiformis f. sp. tritici population has been identified with molecular markers but whether sexual reproduction occurs in China is unknown. In this study, we surveyed barberry plants for infection by rust fungi in the stripe rust “hotspot” regions in Gansu, Sichuan, and Shaanxi provinces; collected barberry plants and inoculated plants of 20 Berberis spp. with germinated teliospores under controlled greenhouse conditions for susceptibility to P. striiformis f. sp. tritici; and tested P. striiformis f. sp. tritici isolates obtained from aecia on naturally infected barberry plants on the wheat genotypes used to differentiate Chinese P. striiformis f. sp. tritici races to determine virulence variations. Different Berberis spp. were widely distributed and most surveyed plants had pycnia and aecia of rust fungi throughout the surveyed regions. In total, 28 Berberis spp. were identified during our study. From 20 Berberis spp. tested with teliospores of P. striiformis f. sp. tritici from wheat plants, 18 species were susceptible under greenhouse conditions. Among 3,703 aecia sampled from barberry plants of three species (Berberis shensiana, B. brachypoda, and B. soulieana) under natural infections in Gansu and Shaanxi provinces, four produced P. striiformis f. sp. tritici uredinia on susceptible wheat ‘Mingxian 169’. Sequence of the internal transcribed spacer (ITS) regions of the four isolates from barberry shared 99% identity with the P. striiformis f. sp. tritici sequences in the National Center for Biotechnology Information database. The four isolates had virulence patterns different from all previously reported races collected from wheat plants. Furthermore, 82 single-uredinium isolates obtained from the four barberry isolates had high virulence diversity rates of 9.0 to 28.1%, indicating that the diverse isolates were produced through sexual reproduction on barberry plants under natural conditions. In addition to P. striiformis f. sp. tritici, sequence analysis of polymerase chain reaction products of the ITS regions and inoculation tests on wheat identified P. graminis (the stem rust pathogen). Our results indicated that P. striiformis f. sp. tritici can infect some Berberis spp. under natural conditions, and the sexual cycle of the fungus may contribute to the diversity of P. striiformis f. sp. tritici in China.


2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Yaqiong Zhao ◽  
Yilin Gu ◽  
Feng Qin ◽  
Xiaolong Li ◽  
Zhanhong Ma ◽  
...  

Stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) is a devastating wheat disease worldwide. Potential application of near-infrared spectroscopy (NIRS) in detection of pathogen amounts in latently Pst-infected wheat leaves was investigated for disease prediction and control. A total of 300 near-infrared spectra were acquired from the Pst-infected leaf samples in an incubation period, and relative contents of Pst DNA in the samples were obtained using duplex TaqMan real-time PCR arrays. Determination models of the relative contents of Pst DNA in the samples were built using quantitative partial least squares (QPLS), support vector regression (SVR), and a method integrated with QPLS and SVR. The results showed that the kQPLS-SVR model built with a ratio of training set to testing set equal to 3 : 1 based on the original spectra, when the number of the randomly selected wavelength points was 700, the number of principal components was 8, and the number of the built QPLS models was 5, was the best. The results indicated that quantitative detection of Pst DNA in leaves in the incubation period could be implemented using NIRS. A novel method for determination of latent infection levels of Pst and early detection of stripe rust was provided.


Plant Disease ◽  
2010 ◽  
Vol 94 (2) ◽  
pp. 221-228 ◽  
Author(s):  
Baotong Wang ◽  
Xiaoping Hu ◽  
Qiang Li ◽  
Baojun Hao ◽  
Bo Zhang ◽  
...  

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici, is a devastating disease in China. Races CYR32 and CYR33 have been predominant in the recent P. striiformis f. sp. tritici population. To develop molecular markers for these races, initially 86 isolates, most of which were collected in 2007 throughout China, were tested on the set of wheat genotypes for differentiating Chinese P. striiformis f. sp. tritici races, and their genomic DNA were amplified with 94 random amplified polymorphic DNA (RAPD) primers. Twelve isolates were identified as CYR33, 14 as CYR32, and 60 as 13 other races. A 320-bp band was identified to be associated with CYR32 with primer S1271 (5′-CTTCTCGGTC-3′), and a 550-bp band was identified to be specific to CYR33 with primer S1304 (5′-AGGAGCGACA-3′). The two bands were cloned and sequenced. Based on the sequences, sequence characterized amplified region (SCAR) markers CYR32sp1/sp2 and CYR33sp1/sp2 were developed to differentiate CYR32 and CYR33, respectively, from other races. The SCAR markers were validated with DNA samples from wheat leaves inoculated with selected isolates from the 86 isolates and urediniospore DNA samples from an additional 63 isolates collected from 2006 to 2009. The detection of CYR32 and CYR33 with the SCAR markers was completely consistent with the results of the race identification with the set of differential wheat genotypes. Thus, the markers are highly reliable for identification of the two races.


Plant Disease ◽  
2007 ◽  
Vol 91 (12) ◽  
pp. 1669-1674 ◽  
Author(s):  
J. Zhao ◽  
X. J. Wang ◽  
C. Q. Chen ◽  
L. L. Huang ◽  
Z. S. Kang

Monitoring the pathogenic fungus of wheat stripe rust, Puccinia striiformis f. sp. tritici, plays a key role in effective control of the disease. In the present study, we developed a specific and sensitive polymerase chain reaction (PCR) assay for detecting the pathogen in wheat (Triticum aestivum) leaves. A pair of primers (PSF and PSR) was designed based on the internal transcribed spacer (ITS) region sequence of P. striiformis f. sp. tritici. PCR products that were amplified with universal primers ITS1 and ITS4 were cloned into pGEM-T Easy vectors and sequenced. The ITS sequence was compared with those of P. striiformis f. sp. tritici, P. triticina, P. graminis f. sp. tritici, Blumeria graminis f. sp. tritici, Fusarium graminearum, Rhizoctonia cerealis, and Bipolaris sorokiniana, which are associated with early symptoms of foliar diseases on wheat. Specificity of the primers was tested in the PCR assays using DNA extracted from all tested P. striiformis f. sp. tritici isolates, other fungal species, and healthy and infected wheat leaves sampled around stripe rust foci in wheat fields, different days after inoculation with P. striiformis f. sp. tritici, as well as asymptomatic wheat leaves sampled around stripe rust foci in the fields. A PCR product of 169 bp was amplified from DNA of all P. striiformis f. sp. tritici isolates. The primers did not amplify DNA from the other tested fungal species. The pathogen was detected from asymptomatic wheat leaves inoculated with P. striiformis f. sp. tritici under greenhouse conditions, as well as leaves sampled around stripe rust foci in wheat fields. Under optimum conditions, the PCR assay was highly sensitive and required only 0.1 pg of the target DNA for a detectable and reliable amplification with the PSF and PSR primers.


2020 ◽  
pp. PHYTO-05-20-017
Author(s):  
Yuheng Yang ◽  
Fengfeng Zhang ◽  
Tianyu Zhou ◽  
Anfei Fang ◽  
Yang Yu ◽  
...  

Plant subtilases (SBTs) or subtilisin-like proteases comprise a very diverse family of serine peptidases that participates in a broad spectrum of biological functions. Despite increasing evidence for roles of SBTs in plant immunity in recent years, little is known about wheat (Triticum aestivum) SBTs (TaSBTs). Here, we identified 255 TaSBT genes from bread wheat using the latest version 2.0 of the reference genome sequence. The SBT family can be grouped into five clades, from TaSBT1 to TaSBT5, based on a phylogenetic tree constructed with deduced protein sequences. In silico protein-domain analysis revealed the existence of considerable sequence diversification of the TaSBT family which, together with the local clustered gene distribution, suggests that TaSBT genes have undergone extensive functional diversification. Among those TaSBT genes whose expression was altered by biotic factors, TaSBT1.7 was found to be induced in wheat leaves by chitin and flg22 elicitors, as well as six examined pathogens, implying a role for TaSBT1.7 in plant defense. Transient overexpression of TaSBT1.7 in Nicotiana benthamiana leaves resulted in necrotic cell death. Moreover, knocking down TaSBT1.7 in wheat using barley stripe mosaic virus-induced gene silencing compromised the hypersensitive response and resistance against Puccinia striiformis f. sp. tritici, the causal agent of wheat stripe rust. Taken together, this study defined the full complement of wheat SBT genes and provided evidence for a positive role of one particular member, TaSBT1.7, in the incompatible interaction between wheat and a stripe rust pathogen.


2016 ◽  
Vol 2016 ◽  
pp. 1-7
Author(s):  
Juan Wang ◽  
Yang Wang ◽  
Xinjie Liu ◽  
Yuanliu Xu ◽  
Qing Ma

The plant cytoskeleton, including microtubules and microfilaments, is one of the important factors in determining the polarity of cell division and growth, as well as the interaction of plants with invading pathogens. In defense responses of wheat against the stripe rust (Puccinia striiformisf. sp.tritici) infection, hypersensitive response is the most crucial event to prevent the spread of pathogens. In order to reveal the effect of microtubules on the hypersensitive cell death and H2O2accumulation in the interaction of wheat (Triticum aestivum) cv. Suwon 11 with an incompatible race, CYR23, wheat leaves were treated with microtubule inhibitor, oryzalin, before inoculation. The results showed that the frequency of infection sites with hypersensitive response occurrence was significantly reduced, and hypersensitive cell death in wheat leaves was suppressed compared to the control. In addition, the frequency and the incidence of infected cells with H2O2accumulation were also reduced after the treatment with oryzalin. Those results indicated that microtubules are related to hypersensitive response and H2O2accumulation in wheat induced by the stripe rust, and depolymerization of microtubules reduces the resistance of plants to pathogen infection in incompatible interaction, suggesting that microtubules play a potential role in the expression of resistance of wheat against the stripe rust fungus.


Plant Disease ◽  
2020 ◽  
Vol 104 (9) ◽  
pp. 2434-2439 ◽  
Author(s):  
Furong Peng ◽  
Miu Si ◽  
Yang Zizhu ◽  
Yuhang Fu ◽  
Yuheng Yang ◽  
...  

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important and devastating diseases of wheat; therefore, it is necessary to rapidly and accurately quantify fungicide effectiveness to monitor Pst sensitivity and manage the disease. In this study, a rapid method of quantifying the fungicide effectiveness with detached leaves was developed. The results showed that 0.5% water agar containing 75 μg/ml of 6-benzylaminopurine and filter paper worked the best for maintaining wheat leaves. The disease incidences of different concentrations of spore suspension were compared. When the spore concentrations were 5 and 10 mg/ml, the disease incidences had no significant differences at 12 and 15 days after inoculation (P < 0.05). Fungicide treatment tests revealed that there were no significant differences in the efficacies of triadimefon on rust suppression between detached leaves in the culture dishes and direct spray on seedlings. We also developed a Photoshop software method that can replace the current classification method and accurately measure the proportion of sporulation area on infected leaves. The sensitivity baseline of Pst to triadimefon was estimated as 0.1453 ± 0.0081 μg/ml, and all the values of EC50 were tested for normal distribution using the Shapiro–Wilk test (W = 0.204). The baseline can be used to test the sensitivity of different Pst isolates to triadimefon.


Sensors ◽  
2020 ◽  
Vol 20 (16) ◽  
pp. 4550
Author(s):  
Huajian Liu ◽  
Brooke Bruning ◽  
Trevor Garnett ◽  
Bettina Berger

The accurate and high throughput quantification of nitrogen (N) content in wheat using non-destructive methods is an important step towards identifying wheat lines with high nitrogen use efficiency and informing agronomic management practices. Among various plant phenotyping methods, hyperspectral sensing has shown promise in providing accurate measurements in a fast and non-destructive manner. Past applications have utilised non-imaging instruments, such as spectrometers, while more recent approaches have expanded to hyperspectral cameras operating in different wavelength ranges and at various spectral resolutions. However, despite the success of previous hyperspectral applications, some important research questions regarding hyperspectral sensors with different wavelength centres and bandwidths remain unanswered, limiting wide application of this technology. This study evaluated the capability of hyperspectral imaging and non-imaging sensors to estimate N content in wheat leaves by comparing three hyperspectral cameras and a non-imaging spectrometer. This study answered the following questions: (1) How do hyperspectral sensors with different system setups perform when conducting proximal sensing of N in wheat leaves and what aspects have to be considered for optimal results? (2) What types of photonic detectors are most sensitive to N in wheat leaves? (3) How do the spectral resolutions of different instruments affect N measurement in wheat leaves? (4) What are the key-wavelengths with the highest correlation to N in wheat? Our study demonstrated that hyperspectral imaging systems with satisfactory system setups can be used to conduct proximal sensing of N content in wheat with sufficient accuracy. The proposed approach could reduce the need for chemical analysis of leaf tissue and lead to high-throughput estimation of N in wheat. The methodologies here could also be validated on other plants with different characteristics. The results can provide a reference for users wishing to measure N content at either plant- or leaf-scales using hyperspectral sensors.


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