scholarly journals Gene Expression Evaluation of Multigenic Cotton (Gossypium hirsutum) against Cotton Leaf Curl Virus

2021 ◽  
Vol 26 (01) ◽  
pp. 169-176
Author(s):  
Muhammad Rizwan Afzal

One of the most crucial threats limiting the sustainable production of cotton is cotton leaf curl disease (CLCuD). There is dire need to produce a resistant variety that can combat CLCuD. For this purpose, virus resistant transgenic cotton plants (MNH-786) with C4 gene construct at T3 generation were selected and sown. Young fresh leaves of multigenic variety of MNH-786 were collected to confirm the transformed construct. Infected whiteflies were used for spreading on transgenic cotton MNH-786 variety with C4 construct to check percentage of infection. Whiteflies were collected from infected cotton plants showing CLCuD and reared in lab to increase the population of whiteflies. After 15 days of feeding, infected leaves of transgenic plants were collected and total DNA of infected leaves of transgenic cotton plant with virus load was extracted. At maturity, data of morphological characteristic was taken from the transgenic cotton plants of MNH-786 and control plants. Resistant transgenic cotton plants showed < 0.5% disease index and recorded more plant height in field condition. Total number of bolls per plant was 20% more in tolerant plants and 40% more in resistant plants as compared to susceptible plants. Molecular analysis of transgenic plants showed clear evidence that expression of construct 4 virus resistant gene against begomoviruses in resistant and tolerant group of transgenic plants was more as compared to susceptible group and control. © 2021 Friends Science Publishers

2016 ◽  
Vol 23 (3) ◽  
pp. 358-362 ◽  
Author(s):  
Sayed Sartaj Sohrab ◽  
Mohammad A. Kamal ◽  
Abdul Ilah ◽  
Azamal Husen ◽  
P.S. Bhattacharya ◽  
...  

2020 ◽  
Vol 56 (No. 3) ◽  
pp. 154-162
Author(s):  
Sehrish Khan ◽  
Muhammad Mahmood ◽  
Sajjad Rahman ◽  
Farzana Rizvi ◽  
Aftab Ahmad

Over the last decade, the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) CRISPR/Cas9 system has been used by biologists in various fields. In plant biology, the technology is being utilised to manufacture transgenic plants resistant to different diseases. In Pakistan, the Cotton leaf curl virus (CLCuV) (a begomovirus) affects cotton plants causing significant loss to the economy of this agriculturally based country. In the present study, we use the CRISPR/Cas9 system in Nicotiana benthamiana Domin (a model plant) to develop resistance against CLCuV. An interesting facet of the study was the comparison of two constructs (pHSE401 and pKSE401) with regards to their efficacy in the virus inhibition. The pKSE401 vector contained a Cas9 nuclease and two guide RNAs (gRNAs), one targeting the Replication associated protein (Rep) gene and the other targeted the βC1 gene of the Betasatellite. The vector pHSE401 had only one sgRNA that targeted the (Rep) gene. Both genes that are intended to be targeted play important roles in the replication of CLCuV. Plants infiltrated with pKSE401 exhibited a delay in the development of the symptoms of the disease and showed lower virus titres. Our proposed multiplexing approach gave efficient results of the resistance in the model plants, and the results in this communication may be extended to the CRISPR/Cas9 based editing of cotton plants.


2016 ◽  
Vol 58 (12) ◽  
pp. 807-820 ◽  
Author(s):  
Aneela Yasmeen ◽  
Sarfraz Kiani ◽  
Afshan Butt ◽  
Abdul Qayyum Rao ◽  
Faheem Akram ◽  
...  

AoB Plants ◽  
2018 ◽  
Author(s):  
Rakhshanda Mushtaq ◽  
Khurram Shahzad ◽  
Shahid Mansoor ◽  
Zahid Hussain Shah ◽  
Hameed Alsamadany ◽  
...  

Plant Disease ◽  
2000 ◽  
Vol 84 (7) ◽  
pp. 809-809 ◽  
Author(s):  
S. Mansoor ◽  
S. Mukhtar ◽  
M. Hussain ◽  
I. Amin ◽  
Y. Zafar ◽  
...  

The current epidemic of cotton leaf curl disease (CLCuD) in Pakistan started in 1988 with the natural host range limited to a few plant species in the family Malvaceae. However, we have observed expansion in the host range of the virus, and several non-Malvaceous plants were found to be infected with the virus. Characteristic symptoms of CLCuD such as leaf curl and enations have been observed on radish plants, primarily in kitchen gardens. However, in 1999, levels of infection of 10 to 90% were observed both in commercial fields and kitchen gardens in the Punjab province of Pakistan. Both symptomatic and nonsymptomatic samples were collected from five different locations. Total DNA was isolated, dot-blotted on nylon membrane, and a full-length clone corresponding to DNA A of cotton leaf curl virus was labeled with 32P dCTP and used as a probe for the detection of a begomovirus. Strong signals were observed in symptomatic plants while no signals were observed in nonsymptomatic plants. Infection with a begomovirus was further confirmed by polymerase chain reaction (PCR) using degenerate primers for DNA A (1). Primers specific for the two distinct begomoviruses associated with CLCuD were also used in PCR reactions (2), and products of the expected size were obtained from all symptomatic samples, confirming infection with begomoviruses similar to those associated with CLCuD. A full-length probe of a nanovirus-like molecule associated with cotton leaf disease (3), called DNA 1 was labeled with 32P dCTP and detected the virus only in symptomatic plants. Similarly, primers specific for DNA 1 (3) amplified a product of expected size when used in PCR. On the basis of symptomatology and the detection of specific viral components associated with the disease, we confirmed that radish plants are infected with Cotton leaf curl virus (CLCuV). Since radish is a short duration crop, infection of CLCuV in radish may not serve as a direct source of infection for the next cotton crop. However, it is a potential threat to tomato crops which overlap with radish in the Punjab province. The detection of CLCuD in radish is another example of the mobilization of begomoviruses to previously unknown hosts. References: (1) M. R. Rojas et al. Plant Dis. 77:340, 1993. (2) S. Mansoor et al. Pak. J. Bot. 31:115, 1999. (3) Mansoor et al. Virology 259:190, 1999.


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