Analyzing Secondary Metabolic Pathways in Plant Cells and Experiments in Active Compound Enrichment

The medicinal material samples and inheritance files sent by the Nara Clan were sorted by Rinto, covering half of the experimental table. He did not rush to design the ripening configuration, but first settled on the core logic of the entire subject: first understand "what it is", and then talk about "how to change it".

It has never been difficult to forcibly accelerate the growth cycle of plants. Even Genin, who has just learned Medical Ninjutsu, can use Yang Release Chakra to stimulate the rapid division of plant cells and keep the seedlings in a few days. It will grow into an adult plant within 10 days. But the difficult thing is how to ensure or even increase the active ingredient content of medicinal materials while accelerating growth - after all, the core value of medicinal materials does not lie in how big the plant grows, but in the secondary metabolites accumulated in it that can cure diseases.

Nara Clan's hundreds of years of failed ripening experiments have also confirmed this point: all medicinal materials that have been forcibly ripened have suffered serious loss of active ingredients, and no matter how tall the plants grow, they are just useless weeds with no medicinal effect.

Therefore, the first core problem that Rinto needs to solve is not to accelerate growth, but to completely dismantle the synthesis path of the active ingredients of medicinal materials, clarify the accumulation rules of secondary metabolites, and ensure that while accelerating growth, the active ingredients can be synchronized or even excessively enriched.

He first selected two of the most core and representative medicinal materials as the first batch of research subjects: one is Dragon Blood Grass, which is matured in three years. Its core active ingredient is dragon blood, which can effectively activate body activity and repair visceral damage. It is the core medicinal material for the treatment of serious injuries in wartime; the other is Ice Lotus, which is matured in five years. Its core active ingredient is Liansinsu, which can effectively inhibit neurotoxins and stabilize mental strength. It is a key medicinal material to detoxify Ninjutsu and soothe the mental collapse of seriously injured patients.

These two kinds of medicinal materials have long growth cycles and complex synthesis paths of active ingredients. They are also the two types with the highest failure rate in the Nara Clan ripening experiment. As long as these two kinds of medicinal materials can be handled, other medicinal materials only need to adapt and adjust the parameters.

Rinto first walked into the sterile operation room and isolated plant cells at different growth stages from fresh samples of dragon blood grass and ice lotus. From the embryonic cells of seeds, to the meristems of seedlings, to the cells of leaves, rhizomes, and medicinal parts of mature plants, they were prepared into samples one by one and placed in petri dishes.

The first step he needs to do is to analyze the entire cycle of cell metabolism pathways. Use the high-power Chakra microscope to track plant cells at different growth stages, from photosynthesis, basal metabolism, to the synthesis, transport, and accumulation of secondary metabolites, and accurately locate the key nodes, key enzymes, and corresponding gene regulation sites for the synthesis of active ingredients.

This is an extremely tedious task that requires extreme patience and precision. The secondary metabolic pathways of plants are extremely complex, and the synthesis of active ingredients involves dozens of biochemical reactions. Each step has corresponding enzymes and regulatory mechanisms. If you are not careful, you will miss key nodes.

But for Rinto, who has been dealing with precise Chakra configurations and parameters to three decimal places all year round, he is already familiar with this kind of precise tracking work. He built a set of micro-monitoring arrays that can track metabolic changes in cells in real time. He placed cell samples at different stages into corresponding monitoring environments and recorded material changes, Chakra characteristics, and metabolic pathways in cells 24 hours a day.

For two whole days and two nights, Rinto barely left the sterile operating room. His eyes were firmly locked on the cells under the microscope, and he kept using the charcoal pencil in his hand to draw the complete synthesis path of dragon blood hormone and lianxin hormone on the slate, marking each key reaction step, rate-limiting step, and corresponding regulatory site.

He found that the dragon's blood of dragon's blood grass is mainly synthesized in the parenchyma cells of the rhizome. The peak period of synthesis is in the second to third year of plant growth. After the basic vegetative growth is completed, a large amount of secondary metabolism will be accumulated. This is also why the forcibly ripened dragon's blood grass will only grow stems and leaves, but has no medicinal effect - the speed of vegetative growth is forcibly accelerated, but the control switch of secondary metabolism is not turned on, and the effective ingredients have no time to be synthesized.

The synthesis pathway of Liensinsu in Ice Lotus is more complex, and it requires slow accumulation in three growth cycles in a low-temperature environment before it can reach a medicinal concentration in the stamens. Conventional ripening not only fails to activate the synthesis of Liensinin, but may even directly inhibit the entire metabolic pathway due to changes in temperature and growth rate.

After clarifying the synthesis path and accumulation rules of active ingredients, the next core is how to break the restrictions of the growth cycle on secondary metabolism and achieve early and efficient enrichment of active ingredients.

Rinto did not use the rough stimulation of Chakra, but designed a set of precise secondary metabolism regulation configurations based on the metabolic pathways that had just been dismantled. The core of this set of configurations is not to forcefully instill something into the plants, but to precisely regulate the metabolic switches of the plant cells themselves:

First, activate the key regulatory sites of secondary metabolism in advance, breaking the inherent rule of "complete vegetative growth first, then start secondary metabolism", so that plants can start the synthesis of active ingredients in the seedling stage;

Second, strengthen the key enzyme activity in the rate-limiting step, increase the synthesis efficiency of active ingredients several times, and solve the problem that the synthesis rate cannot keep up with the growth rate;

Third, optimize the transport and accumulation mechanism of the product so that the synthesized active ingredients can be stably stored in the cells of the medicinal site without being catabolized and continuously enriched.

After the theoretical design was completed, Rinto immediately started verification experiments at the cellular level. He used extremely fine Chakra to precisely introduce the regulatory configuration into the callus cells of Dragon Blood Grass and Ice Lotus, and monitored the synthesis of active ingredients in the cells in real time.

The results of the experiment far exceeded expectations.

It originally took more than two years of growth to initiate the synthesis of a large amount of dragon's blood. In the cells introduced into the regulated configuration, obvious signs of synthesis were detected in just 12 hours. After 72 hours, the concentration of dragon's blood in the cells had reached the level of three-year-old wild dragon's blood.

Liensinsu, which has extremely high environmental requirements and complex synthesis pathways, has successfully started synthesis under the control of its configuration. Even in a normal temperature culture environment, the synthesis efficiency is far higher than that of ice lotus seedlings growing naturally.

More importantly, during the entire process, the growth and division of cells were not affected in any negative way, and the basal metabolism was completely normal, achieving the simultaneous advancement of "vegetative growth" and "secondary metabolism", and even the enrichment efficiency of active ingredients was higher than that of naturally growing plants.

Rinto did not stop there. He conducted multiple sets of optimization experiments to adjust the configuration parameters to further improve the synthesis efficiency of active ingredients while ensuring the stability of the regulation and preventing metabolic disorders. He even optimized the synthesis steps based on the metabolic path of disassembly, reducing the loss of intermediate products, so that the final enrichment of dragon blood element and lianxin element reached more than 1.5 times that of wild medicinal materials.

The verification at the cell level was a great success. Rinto organized and archived all the experimental data, metabolic pathway maps, and configuration control parameters. He finally understood the core difficulty of this topic. Next, he was to solve the second problem: how to forcefully accelerate the overall growth cycle of the plant without destroying the metabolic balance, and compress the three-year growth process to less than three months.

He put down the charcoal, rubbed his sore eyebrows, and looked at the bottle of three-year-old wild dragon's blood sample sent by the Nara Clan in the corner of the experimental table. The natural law of life takes three years to complete, and he wanted to use the precise Chakra configuration to compress it to the extreme.

It was already bright outside the window, and the early morning sunlight shone in through the window and fell on the slate full of formulas and diagrams. Rinto stood up, poured himself a glass of water, took two sips, picked up the charcoal again, laid out a new blank slate, and began to design the forced acceleration configuration of the plant growth cycle.