A leading plant scientist, Dr Nasheman Ashraf, currently Principal Scientist at CSIR-IIM, was the first to create a data bank of 64000 Saffron genes and publish the transcriptome data, something that many others followed later. Her Laboratory in Srinagar is working to find answers to critical questions that prized spice faces in Kashmir: Why it cannot grow beyond Pampore? How to manage bulb rot? Why is the yield so low? In a detailed interview with Masood Hussain and Humaira Nabi, Dr Nasheman revealed that she has identified a microbe that enhances the yield and she is attempting to use tissue culture for making disease-free corms

KASHMIR LIFE (KL): Given the fact that Jammu and Kashmir is an agrarian state, how important is the study of plant science for us?

DR NASHEMAN ASHRAF (NA): The economy of Jammu and Kashmir is predominantly dependent on agriculture and horticulture. Besides, Kashmir Himalayas are home to a wide range of medicinal plants. On that account, the importance of plant science in the region is far-reaching. This puts a lot of responsibility on our scientists to attain as much knowledge as they can about the subject and provide solutions to the various problems faced by the farmers, such as global warming and climate change.  It is also important to be very well-versed with all the new techniques and technology of plant science so that we can disseminate that information to the people. All this will contribute significantly to the expansion of our economy.

KL: How you got into the plant sciences? What was your academic journey?

NA: My primary education was completed at DAV Magarmal Bagh. I graduated from SKUAST’s Faculty of Agriculture, Wadura. I passed the Indian Council of Agricultural Research (ICAR) postgraduate admission examination with an All India Ranking (AIR) of 25. I attended GB Pant University of Agriculture and Technology, Uttarakhand and completed my post-graduation in Biochemistry. During my master’s, I passed all competitive tests, including NET, GATE, and CSIR-JRF.

Following that, I earned my doctorate from The National Institute of Plant Genome Research in New Delhi. I was able to select my own mentor because I topped the PhD admission examination. I chose one of the female researchers, who proved to be a wonderful guide and helped me get the most out of my research. The work culture that I was subjected to there is the best that I have seen so far. And I really dream of witnessing the same here in Kashmir one day, so that we could make the best out of our abilities.

KL: You were born and raised in an urban setting. What incited you to study agriculture, essentially a peripheral phenomenon?

NA: No matter how modern we become, one can never shrug off the importance of agriculture for the progress of our economy. With this realization, I wrote the entrance examination conducted by SKAUST-K. At the Faculty of Agriculture Wadura, we were blessed to have the best teaching faculty, who inculcated the curiosity of research in us at a very young age.

KL: What was the focus of your PhD research and what were the major takeaways?

NA: My research was based on chickpeas. We studied Fusarium wilt- a widespread plant disease in chickpeas that impacts its yield. We did transcript profiling of susceptible and resistant genotypes during chickpea-Fusarium interaction. The study led to the identification of a set of differentially expressed genes, among which some were common to both the genotypes while a subset of genes was specific to either of the genotype. Based on this data we developed pathways which make a species susceptible and another resistant to the pathogen. The chickpea was used as a model plant and our research can be applied to many other related plants.

KL: What followed your PhD programme?

NA: It took me six years to complete my PhD. Only a month after I defended my PhD, I was offered a scientist fellow position at the Indian Institute of Integrative Medicine (IIIM), Jammu. In 2013, I shifted to the IIIM branch laboratory in Srinagar. In 2015, I was awarded CSIR Raman Research Fellowship to work as a visiting scientist at the University of Kentucky, USA.  In 2018, I was awarded an international fellowship by European Molecular Biology Organization (EMBO) to work in Spain. This year I was awarded the SERB POWER fellowship by SERB, Government of India. This fellowship is given to outstanding women researchers working in India.

KL: For good research, you need to ask very good questions. So, what were the questions that you tried to answer in USA and Spain?

 NA: The mandate of IIIM is drug discovery from Natural Products (both of plant and microbial origin). When I joined IIIM, I started working on Crocus sativus commonly known as saffron because of its great significance in Kashmir’s agricultural economy. I chose the crop under the ethos of helping society in terms of my research work.

Saffron is the most expensive spice in the world due to its organoleptic properties (it imparts colour, flavour and aroma). Saffron also has huge pharmaceutical potential. The latest study from IIIM has revealed saffron to possess the ability to delay Alzheimer’s disease and it is also used in the treatment of depression. Saffron is also known for its anti-cancer activities.

The colour and flavour imparting compounds are synthesized in the stigma of the flower in Crocus sativus. Hence the dried and desiccated stigmas of Crocus sativus from the saffron. There are a few other species of Crocus where these compounds are synthesized in other flower parts also. But in Kashmir, we only have Crocus sativus.

I worked on biosynthetic pathways which synthesize saffron compounds and the mechanism that regulates this pathway. We can use this information to manipulate the pathway of Crocus for enhanced apocarotenoid content and hence better quality saffron. We require the cultivation of 2 – 2.5 lakh flowers for one kg of saffron. Our production rate is unable to meet consumer demands. The process, therefore, is very important for making the production of saffron more lucrative.

While I was serving my EMBO fellowship in Spain, they were already working on Saffron there. In fact, the laboratory I worked in is the one where most of the molecular biology work on saffron has been done. It helped me to incorporate their expertise in my research. I got to study some of the plant samples, which produce saffron in petals as well. We worked on the genes which control tissue-specific production of Crocus apocarotenoids.

 KL: Saffron has long been associated with our culture. However, it is solely cultivated in Pampore. Don’t other parts of Kashmir have the capacity to produce saffron?

NA: Since I joined IIIM, I have been growing saffron in our experimental field located in Srinagar.

I am currently working on a project which is sponsored by the department of biotechnology, Government of India, in which we experiment with the extension of saffron cultivation in non-traditional areas of Kashmir. Last year, I distributed saffron bulbs among a group of my students belonging to various districts for cultivation. We covered Kashmir’s all 10 districts and found that it grew in all the 10 districts.

Last year we had a very good result but the data from this year, which is yet to be collected, is very crucial. We will be collecting saffron samples from all the 10 districts, which will be used for quantifying various components like crocin, picrocrocin and safranal using HPLC. These components are responsible for imparting colour and fragrance to saffron.  The conclusion of this study will answer if any other area of Kashmir has the potential for cultivation of saffron of quality at par with the one produced in Pampore.

KL: What are the indications of the study so far?

NA: I think yes we can grow Saffron in other parts of Kashmir as well. However, we must be careful to ensure that it is either cultivated on raised beds or in karewa. Saffron needs relatively little water, and the bulb will decay if there is waterlogging.

KL: Of the 10 districts, which one do you think has the potential of growing saffron better?

NA: It is too early to comment because we had procured corms from Pampore only and cultivated them at other places. Now that the corms are in the soil at different places in different districts for one year, data collected this year will clear all the doubts.

We will be sharing the statistics with the government, which will encourage people to cultivate saffron in non-traditional areas, which we find promising. We have a vast land area available, unlike Spain or Iran which grows Saffron in vertical installations because of less land availability. So, we will try to make the best out of our resources.

Dr Nasheeman Ashraf, a prominent Kashmir scholar, is a principal scientist at CSRIR-IIIM and working on the world’s costliest spice, the saffron. KL Image: Hilal Shah

KL: What makes the saffron produced in Kashmir more expensive than that of Iran or Spain?

NA: The genome of saffron is the same throughout and there is no chance of genetic variation (only by natural mutations). Vegetative propagation takes place in saffron, which makes the offspring a clone of the parent as they are identical in their genotype. So, genetically we cannot say that the saffron grown here is any better than the rest of the world. However, micro-climatic conditions might have some impact on the quality of saffron.

KL: Science comparatively is a difficult subject. You might work on a project for years and not get an expected outcome. What are the challenges that you face here as a female scientist?

NA: When I first started research on saffron, I had to start from scratch. My main objective was genetic engineering of Crocus and for this, we need to know about the genes involved in a biological pathway. Back then only a few genes were known from saffron. So I had to start by developing a gene database which consisted of around 64000 genes. We were the first to publish transcriptome data of saffron; many other laboratories across the world followed us. We did a comprehensive transcriptome analysis of Crocus for the discovery and expression of genes involved in apocarotenoid biosynthesis. This whole job was challenging. For this work, I had to collaborate with other people working in other institutes and this demanded travel outside the state which for a woman was often full of difficulties.

I was working in one of the best laboratories with state-of-the-art infrastructure but I decided to come back and serve my people. However, I was baffled when I was given a single room with not even a single piece of equipment at all. I remember borrowing a pipette set from one of my colleagues in Jammu. I wrote projects, and received grants and with that, I brought equipment for my laboratory. I had to frequently commute between Jammu and Delhi for experiments to have an access to the required equipment.

So, it is definitely very challenging and coupled with the traditional stereotypical work culture makes it more difficult. Being a woman you have to be home early in the evening. And in Kashmir, we do not have the culture to work late in laboratories and offices. So managing all the work in fixed timelines was many a time, draining. In the rest of the world, scientists work day and night because we have a passion for experiments and research. So, we as a community must respect the discrete time frame of scientists especially women who might even have to work on Sundays and other holidays.

KL: What kind of research are you currently doing on saffron and how is that going to help the saffron farming community?

 NA: We started with basic research on saffron we developed a gene database, and identified a few genes which are involved in the biosynthetic pathway which produces saffron compounds (these compounds are called apocarotenoids). We also identified some novel genes which control or regulate the production of these apocarotenoids. The work has been published in reputed international journals like Plant Molecular Biology, BMC Genomics, Physiolgia Plantarum etc. We also identified a gene which imparts drought stress tolerance to Crocus. This was a very good study and was published in a highly reputed journal, the Journal of Biological Chemistry.

Now we are trying to apply these findings to improve saffron production and quality. Our group surveyed all the saffron-growing areas in Jammu and Kashmir, like Pampore, Budgam district and Kishtwar. We interacted with farmers and found that there are three major problems which are responsible for the decline in saffron cultivation and production. These include lack of quality planting material (corms), lack of irrigation facility and corm rot disease which infects saffron fields.

We are trying to provide science-based solutions to these problems. Like in our laboratory, we have established a protocol for the development of Crocus corms in vitro i.e., through tissue culture. We are trying to scale up this process so that good quality and disease-free corms can be made available to farmers.

We identified an endophyte (beneficial microbe) from Crocus corms which enhances the production of saffron compounds and also reduced corm rot disease by 50 per cent. This study was published in Nature Scientific Reports. Currently, we are in the process of developing a bio-control agent based on this endophyte.

And as I mentioned before that we have identified a gene which imparts drought tolerance to Crocus, we are trying to use this gene for genetic engineering of Crocus to produce drought tolerant variety. If we succeed, it would be a breakthrough. All these approaches will ensure better production and quality of saffron.

I would like to add that under the SERB POWER fellowship grant I will be working towards metabolic engineering of Crocus. In this project, the biosynthetic pathway of Crocus would be engineered or manipulated in such a way that the metabolic flux shifts towards the production of apocarotenoids. This will help to produce better quality saffron with higher apocarotenoid content.

The only bottleneck for these approaches is that it is very difficult to transform saffron. We are putting all our efforts to establish a transformation protocol for saffron. Once done, we can manipulate pathways to enhance saffron quality.

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