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Eco-culture a New Simple Technique for Clonal Multiplication of Plants


[Plants experimented are Cashew and Coffee]


By John Paily


[This technique emerged from, my interaction with nature and logical thinking, during my stint as a biotechnologist]


Preface A clone is a duplicate of living system and its information. The normal process by which life sustains it self is through mixing of information and formation of new seeds against changing time. The information and characteristic of the plants obtained from seed changes because of mixing of information. This is a serious disadvantage in planning an quantitative economy of agro industry. Humankind has always struggled to predict, plan and reap. He has struggled to overcome uncertainties. Mixing of information and the uncertainty associated with productivity of plants derived from seed was always a concern to him and a clone that over steps the mixing of the information is a solution. Plants are known to be toti-potent. In other words, they retained the capacity to develop complete plants from the parts. It was noted from time immemorial that many parts of plants such as fallen branches twigs, in appropriate condition, could produce a complete plants. This observation gave rise to techniques of rooting the cuttings to get cloned plants.  


Developments in Plant science showed that the plant growth and its differentiation of root and shoot are controlled by ratio of hormones, auxins and cytokinins. This in turn led to experiments to manipulate plant parts in test tubes in the laboratory, in agar mediums, containing artificial nutrients. This research led to inducing and controlling the shoot and root developments from plant parts in Vitro. The mind of human seeking application and exploitation then paved way for a branch of biotechnology called tissue culture industry. The industry called tissue culture and biotechnology in general has failed to grow up to the potentials it proposed in spite of consuming huge resources in terms of research and developments.


The problem faced by the Tissue Culture industry


1] Our knowledge of life and its information process is incomplete. No body knows why nature mixes the information and why plants are toti-potent but not animals and humans.

2] There is no single rule and method applicable to all plants. This makes it mandatory that one develop a method for every species.  This involved huge research input into developing appropriate media, choosing or inventing appropriate hormones [chemical conditions], fixing physical conditions etc. My own experience of observing over 400000 cultures in vitro and life in nature tells me that within the species the plants and animals have individuality. They are unique in space and time. They are the product of energy interaction of the system with energy of the surrounding, which is changing with time. This makes the control of genetic information further difficult. A process developed for one species and variety have tendency to fail when applied to another variety in the same species. Even the physico-chemical factors to which mother plant are exposed also plays an important role. This means a developing a stable technology that maintains the stability of information is rather arduous task.


3] Plants obtained by callus culture [where cells are dedifferentiated and differentiated  to form embryoids which then give rise to plants]   showed higher diversity than the seed derived plants thus defeating the objective of industry oriented research. It exposed the lack of our basic understanding of information and its expression in biological system. There was stability of and uniformity in respect to chosen characteristics in plants obtained from axillary bud multiplication. Commercial multiplication through axillary bud multiplication in plants has been achieved in banana, cardamom. Here too reports exist of unexplained failure and uncertainty. These problems are further amplified by the arduous task of acclimatizing the fragile plants grown in test tube in artificial sterile medium to natural conditions. At the practical level, it included educating and supporting the farmers to handle these delicate plants to its ultimate end. The sum total when truly evaluated is more negative than positive and tissue culture technique has failed to grow up to replace the traditional technique


However, the research, industry and concepts have survived by the magic of modern economy and research, which hides the negative aspect over exaggerates and advertises to create a vortex to lure more investment to keep it alive. This is done against once own consciousness making it more and more fragile. Yet I must admit that with in this unreal growth a space also develops for real growth, which from time to time emerges to support the system.    



The Logic of Eco-Culture concept


The truths that the cells and tissues in test tubes were speaking on my face were contrary to what I was taught and the objective of my research. This led me to free my mind from what is taught and observe living system in nature as a whole from a point of freedom. This interaction with nature from a point of freedom began to open many new paths to my objective that appeared simple and sensible. Eco-culture is one of them.


What every bio-technologist and tissue culturist does, is to mimic what life does instinctively against time and change that goes in cycles. [However, he is ignorant why life does it]. The tissue culture research and industry is built on the realization that the ratio of auxin /cytokinin [hormones] controls the growth and differentiation of plants. Auxin favoring root formation and cytokinins favoring shoot growth. A tissue culturist tying to clone plants tries to develop the physico-chemical condition appropriate to plants growth and then use available cytokinins and auxins to control the growth and differentiation. Observation of the plant system in nature tells us a fact that the plant grows producing differentiation and growth in response to nature and its cycle of energy change. Two observations gave way to eco culture


1] It was noted that most young branches with few leafs when kept in highly humid conditions has potentials to develop roots.


2] The axillary buds are induced to grow when the climatic conditions change. They seemed to react to energy changes in nature. This opened up a possibility of producing cloned plants by controlling the internal mechanism producing the auxin and cytokinins  by simply manipulating the external environment.


The Technique was experimented with cashew and coffee plants. The following are the broad steps involved in it


1] Rooted plants of cashew and coffee were grown in controlled environment to promote the apical vegetative growth, when the plants are gown 4 to 10 steps. The apical meristem along with two leaves was removed and was transferred to another pot for rooting.

2] The decapitated plant is subjected higher energy state [light and nutrients] to initiate the axillary buds

3] The plants were then shifted back to lesser light and over cast conditions; this promotes the growth of the axillary shoot. It is returned to optimal condition for growth. As the axillary shoot grow by two-step of leaves, its apical meristem along with two leaves and a short stem [one-step] is removed and potted for rooting conditions

4] The mother plant is subjected to increased energy conditions [light and nutrients] to produce more axillary shoots.




Note the shoots forming from every leaf axil of plant



    The work was extended and tested with another commercially important plant Coffee





Eco Culture - Coffee Plants

Note the shoots formed from axillary bud


The process can produce linearly increasing number of plantlets and could be stabilized to produce of clonal plantlets continuously and very economically.


In yet another innovative thinking and experimentation when growth of axillary buds of cashew in tissue culture  was not satisfactory or failed to grow beyond a point, pointing  to some limitations in the medium and hormone [physico-chemical factors given to it], led the author to experiment with axillary bud along with a leaf. See Photo below]. The argument was simple the leaf is the production center of food and hormones to the plant. This meant I can use this machine to make good for any fine chemicals and conditions I am failing to give through the medium.


The results obtained showed marginally increased  growth in the shoot. However, it suggested that the need is not fulfilled. This was then combined with another fine observation, that in nature plants respond to climatic changes by producing shoot and root growth. The intensity needed to trigger growth differs from species to species. Cashew was perceived to be at higher end. This prompted the author to experiment it directly in glass house conditions placing the explants in plastic glasses filled with sand. Sun light seemed trigger the mechanism of the leaf, which failed in artificial light conditions of the laboratory. The results was surprising, nearly 90% of the explants not only produced the shoot but also roots, making the whole technology highly feasible.



Axillary Bud with leaf in Culture



This technology, conceptually could be extended to any plants heralding a simple universal cloning technique  that is practicable, economical and has little risk of changing the genetic aspect of the mother stock. 



The concept has the following advantage

1] It is universal.  It can be explored and developed for all plants.

2] The development of technique and standardizing it is cost effective. It needs very little effort from a researcher to develop and refine the optimal energy [physico-chemical] conditions of growth and the quantum changes that initiates the differentiation.

3] Since the plants system is not subjected to artificial medium and hormone, the problem of uncertainty does not emerge. The genetic stability of the mother stock is maintained. It should be noted that all biological system struggles constantly to survive. When plant system is subjected to artificial medium and hormones, the information system is heavily stressed and makes many random changes in order to survive against the stress force acting on it. This is the principle cause for the uncertainty in tissue-cultured plants. The concept eco-culture over comes this aspect.

4] Since the plants are produced in natural condition, it needs no acclimatization.

5] The production cost of the cloned plants is largely reduced.

6] It is practicable in a small scale as well as large scale. The expensive glass house can be replaced with, polythene bags to cover and maintain the humidity


Important Notes

1] The importance of day and night energy cycle and the climatic cycle of nature to living system caught my attention through this innovative thinking. This later formed the foundation to visualize the fundamental design, principle and the process of quantum energy exchange leading to perpetual motion.

2] The work was done nearly two decades back. Author chose not  to release this simple technology because, as researcher and scientist, author could not find any answer why nature mixes its information. Except for the economic potential and quantitative production, author could not find answer to long range impact of hindering the natural process of life. Observation in nature showed that monoclonal plantations have high risk of quick disease spreading. [This will be explained later]. In such situation author decided not to release the technique or develop it as a technology to make money but concentrated on the fundamentals guiding nature

3] One might ask why I am releasing it now - The answer is simple. From the light of knowledge that author gained by looking beyond the genes, knowing the foundation of plant life, having the answer why plant is toti-potent, author sees positive side to cloning plants. These aspect will be discussed as time permits - Read article "Beyond Genes" and "Secret of Consciousness" to know the fundamental difference between plant life, animal life, and human life


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