Introduction 

Tissue culture is a buzzword among all plant-lovers and agriculturists, representing one of the most advanced technologies not only for growing healthy, disease-free plants but also for their conservation and multiplication to meet increasing demands.

This revolutionary process has transformed how we propagate plants, offering a multitude of benefits for agriculture, research, and even the houseplants we cherish.

This month, I had the privilege of speaking with Shariq Hassan, a leading expert in the field of tissue culture and the founder of PlantLife Biotech. Shariq brings a wealth of knowledge and experience to the table, having played a significant role in advancing this innovative technique and building his tissue culture business.

In November 2023, Shariq founded Plantlife Biotech, a company specializing in the production of banana plants, particularly the Grand Naine and Williams varieties. Within a short timeframe, they've achieved a remarkable feat: establishing a commercially operational plant tissue culture lab capable of producing up to 1 million plants annually.

To give you an overview, the topics covered during the interview are:

• Shariq’s Experience and Work in Tissue Culture
• Tissue Culture For Beginners
• Tissue Culture Challenges and Potential Solutions
• Future of Plant Tissue Culture
• View on PCT Services

Shariq’s Experience in Tissue Culture

Anjali: Can you tell us about your journey into plant tissue culture? What initially sparked your interest in this field, and what continues to excite you about it?

Shariq: My early exposure to plant tissue culture came through my father, a Professor of Biotechnology specializing in the field. 

He established one of the first research Plant Tissue Culture Labs in Pakistan back in 1999. As a child, I have many memories of visiting his lab and meeting his students.

So, my exposure to what Plant Tissue Culture is, was something that I got when I was only 9 years old. Seeing the plants growing in jars really was an amusing sight. My father later helped multiple individuals with the R&D of setting up their own Plant Tissue Culture lab. These labs have been doing great business. That’s when it clicked me, to utilize our in-house R&D, i.e. my father, and set our own lab as well.

Anjali: What plant are you working on currently? Why do you prefer only this plant over the others? What did you like most about it?

Shariq:  At this point of time, we are working on Banana Plants, while also side by side optimizing protocols for Date Palm. 

For any business to run sustainability there should be a business case for it. As I mentioned, we have helped other players to establish their own lab, and all those labs are working on Banana Plants. So there is already a certain level of understanding amongst the farmers of how PTC plants can help their business. 

But, we have a second mover advantage here, with the in-house R&D, and already a surging demand it makes a great business case for us to work on Banana. 

What I liked most about working on Banana in comparison to Date palm is the short growing cycle. In just 20-25 days one can easily sub-culture the Banana cultures whereas in Date palm, the cycle gets quite long. Moreover, my province Sindh grows almost 90% of the total banana produced in Pakistan so the proximity of target market also played a huge role in our decision.

Anjali: What’s the major barrier that you faced while starting your tissue culture business?

Shariq: Running a PTC lab, is quite capital intensive. Making sure that we are working on a budget and optimizing our resources has been a key. We have focused since the start on waiting for the raw material such as preservatives, hormones, MS Media against import, to start our work. [It] takes time, so our supply chain planning, coupled with in-house R&D and production of major raw material, has helped us to optimize our resources.

Anjali: What problem do people majorly face while growing the Banana plant using the PTC technique? What solution do you propose for the same?

Shariq: I think the most difficult part for us was optimizing the protocol to initiate the culture. The mother plant stock that you have got in the field  needs to go through several layers of aseptic cleaning. Optimization of the protocol is the toughest part for anyone who will be starting of the PTC. 

I would recommend connecting with like-minded people around the world. Write emails and approach via Linked-In. Read multiple research papers for the issue you are trying to resolve and eventually what you seek will find you-Rumi.

Anjali: What aspects of tissue culture do you believe require learning from an expert, rather than being achievable independently?

Shariq: Apart from waiting impatiently for your plants to grow (pun intended) learn everything from an expert. 

Don’t waste your time and resources experimenting directionless. You should know the ins and outs of this skill. I would highly recommend learning from an expert. The protocol of the plant you are working on, maintaining aseptic environment, etc. requires an expert advice.

Beginner Level Questions

Anjali: In a nutshell, what is plant tissue culture, and how does it differ from traditional plant propagation methods?

Shariq: Plant tissue culture is a method of growing plants in sterile conditions using nutrient-rich media. In our case MS MEDIA, enabling the rapid production of large numbers of genetically identical plants. 

Unlike traditional propagation, which relies on natural processes like seed germination or cuttings and is less controlled, tissue culture ensures a sterile environment, precise control over growth conditions, and uniformity of plants. This technique is highly efficient and reduces the risk of diseases, making it a superior alternative for producing high-quality plant clones.

Anjali: What plants would be easiest to work with in tissue culture for someone who’s just starting to work in the area?

Shariq: Any plant, as a rule that can be easily propagated vegetatively in nature proves easiest under invitro multiplication. All those plants in which the risk of contamination is the lowest, however ornamental plants can be picked up first. Then someone should move to the fruit bearing plants.

Anjali: Would you have any suggestions or ideas for beginners interested in beginning a career in tissue culture? If so, how can they do that?

Shariq: Read and understand the basic concepts. Then try to find someone resourceful who either runs a lab or teaches Plant Tissue Culturing. Find PTC labs in your area and do internships there. Try to learn from an experienced person before starting your own lab. Even if you don’t have the degree in Biosciences, Biotechnology etc. it is still possible for you to learn PTC.

Anjali: What educational preparation would you recommend to someone interested in making their career in tissue culture?

Shariq: A Bachelor’s and master’s degree in Biosciences would be helpful if anyone wants to make a career in Tissue Culture. Find a professor who has an established lab at university/research organization and work under that professor to gain experience.

Anjali: What resource would you recommend keeping us updated with tissue culture research and news? 

Shariq: I personally recommend LinkedIn and Facebook groups and Plant Cell Technology’s blogs and videos. For LinkedIn, in try to find like-minded people working in the domain of PTC around the world. The amount of information you would learn from the posts and updates shared by your linkedin connections will be exponential. You can also join groups and communities on Facebook, one such group is “Plant Tissue Culture Enthusiasts and Micropropagation” where researchers and commercial labs employees share info with each other and help each other out. 

Anjali: From your experience, what are some of the biggest misconceptions people have about plant tissue culture? 

Shariq:  We have seen the misconception that a Tissue Culture plant is a virus-free plant, this thinking also prevails in commercial tissue culture labs.  Until, and unless, the virus-indexing is not done, and certified initial planting material has not been acquired, there is a chance that through tissue culturing you are multiplying the infected plants. It is highly important to use initial material which has been certified and to check the random plants during acclimatization by ELISA, or PCR based kit to ascertain the status of virus free.

Another misconception we see from the farmer’s side is that maybe Tissue Culturing is an artificial method to produce the plants, however this is not the case.

Technical Questions

Anjali: You mentioned specializing in banana plant production via tissue culture. Can you share some insights or suggestions for other growers cultivating bananas to ensure healthy in vitro growth?

Shariq: Well,

1. First and foremost, make sure that you've got your forecasted supplies in stock
2. Secondly, make sure you have studied the literature of banana micropropagation. You can find multiple research articles online published explaining different stages of Banana Tissue Culturing. 
3. Make sure you are well informed of the kind of media that is required. Along with the hormones, temperature, humidity conditions needed to be given to the cultures at every stage. 
4. You could also go through this research paper for further information. 

Anjali: Can you tell us briefly how you grow Banana in vitro. Like as what explant you use, sterilization method, and media. 

Shariq: Find a good source of initial explant. We prefer having our own mother plant stocks, however there are multiple companies which provide stage 0 cultures. So if you don’t have a mother plant stock then you can purchase stage 0 or 1 cultures and multiply them further.

We take out the meristem from the mother plant. [Then we] pass it through a series of aseptic steps, including washing it with bleach/Clorox multiple times. Finally,. washing it with a contamination control mixture, which we prepare in-house.

We prepare and use standard MS MEDIA however utilize different hormones at different stage of the culturing. For instance, in multiplication stage we use BAP 4-5mg/L, in rooting stage we utilize IBA 1-2mg/L.

Anjali: What factors significantly affect the in vitro propagation of Banana plants?

Shariq: One needs to ensure that the media has been autoclaved appropriately and the tissue culture technician knows from where exactly the callus/TLS needs to be divided from. 

Excessive cuts cause bleeding eventually resulting in making the tissue weak/dead. It’s very important to monitor the temperature, humidity and lightning conditions of the growth room. 

One more thing that is important is to have a decreased contamination rate, to ensure that your plants have nutrients available for consumption. Another important thing one must know [is] the minimum size of explant for subculturing. A plant that is too young can create issues like bleeding, blackening, slow multiplication rate, or ultimately dying.

Advanced

Anjali: What are some of the emerging trends and advancements in plant tissue culture technology?

Shariq: As a relatively young lab, we are always trying to be as agile as possible to adopt the advancements going on in the PTC technology. One very prominent change has been the usage of Bioreactors in the lab, instead of using the jars. This [change] not only helps in reducing the time of sub-culturing but also helps in increasing the efficiency and overall plant multiplication rate and its health. We intend on transforming our lab completely on [to] Bioreactors in the coming cycle. 

Anjali: Based on your experience as a tissue culture expert, what do you consider to be the major limitations in tissue culture that limit its application? 

Shariq: I will be giving my answer from a commercial point of view. So, if the cost of production is high, then eventually the plants you produce will be out of the range of your target market. Hence, cost optimization is very important while running the PTC lab. The first and foremost task of anyone who is working in PTC lab, should be to reduce the contamination rate to at least less than 10%. [Do this] to ensure that not a lot of your plants are wasted due to fungal/bacterial contamination. 

Plants which can be multiplied through cutting method cannot be made feasible by producing them via Tissue Culturing, as the cost in PTC will be more. So choose plants which cannot be easily grown by cutting method. 

Some people have misconception that tissue culture is an unnatural type of propagation and it may not be good for nature, however that is not the case and people need to be educated on it. 

Anjali: How do you think we can overcome those limitations?

Shariq:  Well, for contamination control, one needs to make sure that strict protocol is followed in the lab, at every step. One can also use different products available online such as such as Plant Preservative Mixture (PPM™), or any contamination control mixture to reduce the rate of contamination. 

Anjali: One of this technology's most talked-about limitations is the cost associated with the process. How can this cost be reduced so that the process becomes more affordable? 

Shariq:  Using indigenous raw material helps a lot in reducing the cost of production, we at Plantlife Biotech produce our own raw material such as MS Media, Phytagel and CCM, which is at par with any other brands offering. 

We utilize recycled jam jars for culturing. This work is very energy intensive. [However one can set up their own solar electricity unit to reduce the cost of electricity.

Utilize natural daylight in your growth room by designing it in a way through which maximum sunlight can enter the growth room. 

Rather than doing experiments, try to learn first from an experienced person who is well-versed with the plants that you want to work on, to ensure that both your time and energy are saved. 

Make multiplication more efficient by trying different hormones and optimizing growth room condition, such as humidity, lights, and temperature and reduction in the contamination rate by following strict protocols in the lab.  

Anjali: How do you think tissue culture will revolutionize the agriculture and horticulture areas in the future?

Shariq:  Due to the climate change, there remains an uncertainty. For instance, 2 years ago in Sindh province of Pakistan, due to heavy rainfall there was flooding which destroyed the Banana crop. Now, normally the farmers in Sindh grow Banana through suckers, [but] as the crop was destroyed it was through Tissue Culturing that the farmers were able to get the plants so they didn’t miss the crop cycle. Moreover, as much as this technology gets common, it will be benefiting the farmers.

Virus-indexed plants tend to perform better in the field. Increasing per acre yield for the farmer directly increases their earning, ensuring food-security and poverty alleviation.

I think there will be a very positive impact of this technology on both Agriculture and Horticulture soon. Tissue culture plants can help in reducing the spread of virus in the field can help in quick multiplication of newly released or award-winning variety. Producing earlier hybrid seed by using instant inbred lines. More variegated plants in ornamental industry using soma clonal and other induced variation. Developing novel improved verities using induced and soma clonal variation followed by screening against biotic and abiotic stresses

Anjali: These days, home-based tissue culture labs are trending. What’s your opinion on the same? Do you think it will be effective? What problems can one face while doing tissue culture in such labs?

Shariq: You always start small. Not everyone is blessed to have capital to setup a big lab. However, personally it will be difficult to manage the contamination in a home-based tissue culture lab if clear protocols aren’t established and there isn’t a designated area for every step involved in the process. 

Moreover, economy of scale rule apply make it unfeasible for commercial aspect however for a hobby and learning it is a good thing.

Anjali: Can you discuss the role of cryopreservation in plant tissue culture and its significance for germplasm conservation?

Shariq: As 1cubic cm of callus contains about 10 million cells so potentially 10M plants can be stored in one cubic centimeter. Lots of material, especially old varieties, can be stored in gene bank for future use.

Anjali: As of today, what are some major applications of tissue culture? Where is it mostly being utilized, and for what purpose?

Shariq: Some of the major applications of PTC according to me are:

• Large scale propagation of disease and virus tested uniform materials. 
• Programmed production of planting material. Multiplication of novel varieties like in sugarcane. 
• Producing new varieties in those plants whose donors produced seeds. 
• Genetic engineering for resistant varieties and edible vaccine production.

PCT Product and Services Related Questions

Anjali: How did you find out about us? What do you say about our work in tissue culture, including our products, services, master classes, and education information that we provide through our blogs and videos?

Shariq: I found PCT through your Youtube channel. I think Plantcell Technology is doing a great job in making PTC look cool and attracting the younger generation to take interest in this field. Also, Plantcell Technology’s master classes have made it easier for anyone to attend and learn from experts.

Thank you Shariq for this insightful interview and for sharing your expertise on plant tissue culture with our community.

Shariq’s words echo the exciting potential of this field. If you've ever considered a career in plant tissue culture, now is the perfect time to embark on that journey!

For those interested in getting started, Plant Cell Technology is here to help! Whether you have questions about upcoming classes, essential tissue culture products, or even lab setup, feel free to reach out to us at info@plantcelltechnology.com.

Our team is here to address your concerns and guide you on your path to successful plant culturing.

Happy culturing!