CTAB Protocol for Obtaining High-Quality Total RNA from and Other Related Plants of the Family Malvaceae.

Malvaceae family, also known as the Mallow family, is a family of flowering plants containing and other plants of high medicinal value. This study focuses on the challenges associated with high-quality RNA extraction from and its related plants characterized by high levels of mucilage and phenolic compounds in their tissues. High mucilage and secondary metabolite content pose obstacles in obtaining high-quality RNA, negatively impacting downstream applications, such as gene expression analysis.

Vaccine development: Current trends and technologies.

Despite the effectiveness of vaccination in reducing or eradicating diseases caused by pathogens, there remain certain diseases and emerging infections for which developing effective vaccines is inherently challenging. Additionally, developing vaccines for individuals with compromised immune systems or underlying medical conditions presents significant difficulties. As well as traditional vaccine different methods such as inactivated or live attenuated vaccines, viral vector vaccines, and subunit vaccines, emerging non-viral vaccine technologies, including viral-like particle and nanoparticle vaccines, DNA/RNA vaccines, and rational vaccine design, offer new strategies to address the existing challenges in vaccine development.

Demonstration of seed inoculation of dimer infectious clones of mungbean yellow mosaic India virus-soybean isolate.

Unlabelled: In this study, the full-length components of mungbean yellow mosaic India virus (MYMIV) DNA-A (MW590720 & MW600934) and DNA-B (MW659819 & MW659820) from a soybean isolate were cloned and sequenced. Nucleotide sequence analysis of both MYMIV components revealed > 96% identity and close ancestry with MYMIV isolates from legumes in Southeast Asia. Furthermore, dimeric infectious clones of MYMIV were generated in the pCAMBIA1302 vector, and a seed infiltration protocol was established for mungbean, soybean, and .

First report of apple stem grooving virus infection in loquat from India.

Loquat, commonly known as , is a major subtropical fruit from the family that is native to China but also found in most of Europe and Asia, including India. Apple stem grooving virus (ASGV) infecting loquat was detected using leaf samples collected from Himachal Pradesh (India) through DAC-ELISA followed by RT-PCR assays targeting coat protein (CP), movement protein (MP) and replicase (Rep) regions of ASGV genome. Sequencing of RT-PCR amplicons and sequence analyses revealed that CP, MP and Rep sequences of ASGV loquat Indian isolate of the current study shared a maximum of 98-100% nucleotide sequence identities with the corresponding sequences of available ASGV Indian isolates [LN559078, HE978837, MZ127820, MN912568].

Molecular detection and characterization of (PSTVd) infecting Tomato ( L.) in Karnataka State of India.

Potato Spindle Tuber Viroid (PSTVd) is a non-coding, infectious, small, circular RNA known to cause disease in agricultural and horticultural plants. In the present work, an investigation was conducted in the southern districts of Karnataka state to assess the possible pospiviroid infections on tomato plants that are considered natural hosts for viroids. A total of 83 tomato samples showing disease symptoms (virus or viroid-like) along with healthy ones were collected and subjected to viroid detection by conventional Reverse Transcriptase Polymer Chain Reaction (RT-PCR) using universal (Pospi1-RE/Pospi1-FW) and a specific set of primers (3H1/2H1).

Development of a new Collateral Cleavage-independent CRISPR/Cas12a based easy detection system for plant viruses.

Plant virus spread through various means, from mechanically to the insect vectors and act as obligate parasite, therefore, are extremely challenging to eradicate. Geminiviruses are an important class of viruses which have reported extensively in last two decades on several new hosts. They infect wide range of annual crops and perineal shrubs, therefore, essentially required to detect them on field and dispose to check their vector transmission to healthy crops.

Serological and molecular analysis indicates the presence of distinct viral genotypes of in India.

Unlabelled: Recombination leads to the generation of new viral progeny which remain undetected by routine testing procedures and may be a threat to the infected host. Here, we have characterised the complete genome sequences of two isolates of from apple cv. Red Chief (Palampur) and cv.

Apple scar skin viroid naked RNA is actively transmitted by the whitefly Trialeurodes vaporariorum.

Nucleic acid transfer between plants is a phenomenon which is likely to occur in many ways in nature. We report here the active transmission of Apple scar skin viroid (ASSVd) naked ssRNA species by the whitefly Trialeurodes vaporariorum (Tv). Not only the viroid RNA, its DNA form was also identified from the insect.

A simplified strategy for studying the etiology of viral diseases: Apple stem grooving virus as a case study.

A simple method to amplify infective, complete genomes of single stranded RNA viruses by long distance PCR (LD PCR) from woody plant tissues is described in detail. The present protocol eliminates partial purification of viral particles and the amplification is achieved in three steps: (i) easy preparation of template RNA by incorporating a pre processing step before loading onto the column (ii) reverse transcription by AMV or Superscript reverse transcriptase and (iii) amplification of cDNA by LD PCR using LA or Protoscript Taq DNA polymerase. Incorporation of a preprocessing step helped to isolate consistent quality RNA from recalcitrant woody tissues such as apple, which was critical for efficient amplification of the complete genomes of Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV).