Saionji Biotechnology Center (Shakotan)
Research Project Logbook · Supplementary Report by the Project Leader
Project Number: SF-HK-089-14
Project Name: Germplasm Collection and Evaluation of Cold-Hardy Perennial High-Yielding Forage Grasses
Department: Forage Crops Research Laboratory
Establishment Date: April 10, 1989
Annual Budget: 3.8 million yen
Project Objective: To screen perennial forage grass candidate materials with stable overwintering, high dry matter yield, and good digestibility for the S-Farm beef and dairy cattle feeding system, reducing dependence on purchased hay and imported feed.
Recorder: Toshihiko Yamada
Compilation Date: January 3, 1992
——
Exactly how much is a single blade of grass worth?
If it were three years ago, I would probably tell you that it depends on how many cattle it can feed.
After all, that's what I do.
My name is Toshihiko Yamada. I was born in Sapporo, Hokkaido in 1960. I graduated from the Faculty of Agriculture at Hokkaido University and later spent two years in graduate school studying crop breeding.
After graduating in 1985, I entered a seed company within the region, mainly responsible for comparative trials of forage grass varieties.
I did that job for three years.
To be honest, there were no achievements particularly worth bragging about.
I participated in regional trials for several varieties and collected quite a few local varieties for the company, but there were basically no results that I could actually put on my resume.
Therefore, in October 1988, when the Saionji Biotechnology Center published a recruitment advertisement in the newspaper, I submitted my resume.
The compensation was good.
More importantly, they had their own farm.
People who study crops all know that having a farm and not having a farm are two completely different things.
Previously at the seed company, how many test plots could be arranged in a year depended first on customer orders and whether the farmers were willing to cooperate.
When harvest season arrived, sometimes before we even finished sampling, the farmers were already urging us to clear the land.
Saionji was much more convenient.
They had a farm in Tokachi and a breeding center in Shakotan, with dedicated experimental nursery fields right next to the laboratory, and I heard that the group was also planning to continue expanding its agricultural scale.
During the interview, I asked the person in charge of recruitment whether the company could provide stable experimental fields if the research project required continuous observation for five or six years.
He told me that as long as the research plan was approved, of course it could.
I was very satisfied.
So I came.
At that time, I actually didn't know much about the Saionji Group.
I only knew that it was a zaibatsu enterprise that had suddenly developed in recent years, operating agriculture, food, and logistics, and it seemed to have opened quite a few stores in Tokyo.
As for the eldest young miss of Saionji House, I only learned that such a person existed after hearing colleagues mention it after joining the company.
I heard she was Mr. Shuichi's only daughter, still reading high school in Tokyo.
At that time, almost everything published in the newspapers was about Mr. Shuichi, and he was also the one representing Saionji at various events on television. As for this eldest young miss, not many people outside actually knew much about her.
Of course, I was the same.
In my opinion, she was probably a Kazoku Eldest Miss who would inherit the family business in the future, usually living a life completely different from ordinary people like us.
However, what did that have to do with me?
I was just a researcher who grew grass.
Even inside the Hokkaido research institute, I was just one of many ordinary researchers.
...
Half a year after I came to Shakotan, I finally got my first project independently responsible by myself.
April 10, 1989.
I remember this date very clearly.
Because that day when I signed my name in the project leader column, I was very happy and deliberately wrote the strokes a little more carefully than usual.
The project name was very long.
"Germplasm Collection and Evaluation of Cold-Hardy Perennial High-Yielding Forage Grasses."
Simply put, it was to find a grass with high yield, that wouldn't freeze to death in winter, and that cows were willing to eat.
This matter was very necessary in Hokkaido.
The cows on S-Farm were increasing, and purchased hay and imported feed were not cheap, especially in winter, after forage growth stopped, the entire feeding system had to rely on pre-reserved feed.
Mr. Otsuka hoped we could think of a way.
Ideally, we could find a forage grass suitable for Hokkaido that could be harvested continuously for many years once planted.
This kind of thing didn't sound difficult.
But if it were really that easy, farmers in Hokkaido would have already planted it themselves; where would they still need people like us?
When the office director approved the project, he only gave a budget of 3.8 million yen.
This also included experimental field maintenance, material collection, and analysis consumables.
The money allocated to each project actually wasn't much.
Fortunately, I didn't plan to breed new varieties right from the start.
Collect materials first, then slowly compare them.
So that spring, I led two colleagues and began looking for grass all over Hokkaido.
We dug up rhizomes of wild pampas grass on river beaches, went to hillsides and abandoned pastures to look for plants with better growth, and didn't even spare the wasteland beside the highway.
Whenever we found wild gramineous plants with decent growth, we would stop to take a look. We would dig up a portion of the rhizomes for those we could take away, and collect seeds for those we couldn't.
Among them, the most important ones were miscanthus and silvergrass.
These two plants were very common in Japan, grew fast, and could accumulate a lot of biomass.
The shortcomings were equally obvious.
After growing old, the stems were too hard, and the crude protein content was low, so cows usually didn't like to eat them.
Therefore, the office director had never been very optimistic about this direction.
He said our project was like going to the river beach to pick up treasures; it wouldn't matter if we didn't pick up any, at least we could still accumulate some germplasm resources.
I felt what he said made quite a bit of sense.
It's just that for treasures, since one planned to pick them up, of course one still had to look for them seriously.
...
On June 21, 1989, we went to the lower reaches of the Ishikari River for the third time.
Luck was pretty good that day.
From morning to afternoon, a total of 14 materials were collected, numbered from F-35 to F-48.
Among them, one plant was particularly difficult to dig up.
It grew on the edge of the wetland inside the river embankment, the plant was a bit taller than the surrounding grass, and its underground rhizomes were thick and dense.
Sato held an iron shovel and dug for half a day, but when he exerted force, the shovel handle broke directly.
He was so angry that he sat on the river beach and cursed a few times.
I had no choice but to go over and help.
The two of us tossed and turned for another twenty minutes or so before finally digging out a little rhizome.
We brought it back and weighed it.
1.2 kilograms.
That grass was later designated as F-47.
At the time, we thought it was a silvergrass plant.
After all, the leaves, rhizomes, and growing location were all very similar.
As for whether it had any special features?
Other than being exceptionally difficult to dig up, I couldn't think of a second thing.
Therefore, in that year's records, I only wrote one sentence for it:
"F-47: Collected from the edge of the wetland inside the river embankment in the lower reaches of the Ishikari River, possessing developed underground rhizomes, tentatively classified as Miscanthus genus material."
Next, we went to quite a few places.
By the time collection ended in October, the entire project had registered 236 germplasm materials.
Three plants per material.
All planted in the experimental nursery field.
F-47 was just a very inconspicuous one among them.
In the first year, its growth condition indeed didn't have anything special.
After all, having just been transplanted, most materials were recovering from seedling shock, and no one could expect them to grow exceptionally well right away.
I had originally prepared to observe for at least three years.
Who would have known that in the second summer, the first to discover its uniqueness turned out to be a few deer.
...
July 19, 1990.
That morning, Sato, who was in charge of nursery field management, came to the office to find me and told me that Ezo deer had broken in last night.
My first reaction was a headache.
Because these wild animals didn't understand what experimental blocks were at all, nor did they know that the plants in every plot were numbered.
They only knew to walk wherever there was grass.
Sure enough, when I rushed to the scene, the northern fence already had a large hole, and many plants in the fourth and fifth blocks had their upper halves nibbled away.
Sato was furious.
I wasn't doing much better either.
After all, the materials that had been painstakingly raised for over a year—if they were eaten just like that, this year's experiment would probably have to start all over again.
However, after checking for a while, I found the situation a bit strange.
The deer herd seemed to especially like a few of those materials.
Especially F-47.
The upper stems and leaves of those three plants were almost entirely nibbled clean, while the neighboring F-46 and F-48 only left sporadic bite marks.
At the time, I even thought it was because the location where the deer herd entered happened to be close to F-47.
But the damage situation in other blocks also had a similar tendency.
Some grasses were hardly touched, while others were eaten down to just stems.
Thus, I thought of a question.
Why did the deer particularly like to eat them?
Although wild animals' foraging choices were influenced by many factors and could not be simply attributed to sweetness.
But since this phenomenon had been discovered, it had to be checked.
A few days later, I took a handheld refractometer and re-sampled those nibbled materials.
When the results came out, I even thought the instrument was broken.
The sugar content of F-47's stem sap was 12.4.
Miscanthus materials averaged only 5.1.
Sacchariflorus materials were even lower, only 4.3.
I recalibrated the instrument, changed the sampler, and changed the sampling site.
Measured twice the next day.
12.1.
12.7.
Pretty close.
Sato watched me fussing around from the side and couldn't help saying:
"It seems those deer know how to pick grass better than we do."
I didn't refute him.
Because this time, he might actually be right.
...
Ever since then, I began paying special attention to F-47.
The compositional analysis results in September were even more interesting.
Its dry matter yield, when converted, reached 14.2 tons per hectare, ranking first among all 236 materials.
What surprised me even more was the cellulase digestion rate.
61.3%.
Miscanthus materials only averaged 39.8%.
This meant that under our experimental conditions, the fiber components of F-47 were significantly easier to break down than ordinary Miscanthus.
The subsequently measured lignin content was also on the lower side.
Acid detergent lignin accounted for 5.9% of the dry matter, while the average for Miscanthus materials in the same group was 8.7%.
If you only looked at these indicators, it was simply a rare and valuable material.
Unfortunately, there was another problem.
The crude protein content was only 4.2%.
It was simply too low.
As a high-quality forage grass, it was clearly unqualified.
Moreover, it wasn't long before we discovered a second problem.
In late September, Hokkaido was affected by a typhoon.
After the strong winds passed, we went to the nursery field to inspect.
Most of the other Miscanthus materials could still stand, but a large patch of F-47 had collapsed.
The lodging rate of the three plants was about sixty percent, and there were quite a few broken spots at the base of the stems.
I squatted there and looked at it for a long time.
The benefits of low lignin were now laid out right before my eyes alongside its drawbacks.
Its fibers were easy to decompose, and its stems were also easier to break.
If it falls over whenever there's a strong wind, why would farmers bother planting it?
There was a third problem.
F-47 did not set seeds.
In October, we observed that its inflorescence developed normally, but neither self-pollination nor open pollination yielded plump seeds.
It wasn't until the chromosome counts came out in November that things became clear.
2n = 57.
Triploid.
Judging by its morphological characteristics, F-47 was very likely a naturally formed hybrid individual between Miscanthus and Sacchariflorus.
Due to abnormal chromosome pairing, it had almost no ability to set seeds normally.
In other words, it could only reproduce through underground rhizome division, or perhaps try tissue culture in the future.
I finally understood why no materials particularly similar to it were found on that river beach of the Ishikari River.
Such natural hybrid individuals were not easy to appear in the first place.
Even if they did appear, they might not be able to reproduce stably.
As for how many years F-47 had actually grown there, and whether there were other identical individuals around, I didn't know.
If that shovel hadn't broken back then, if Sato had found it too hard to dig and decided to switch to another plant...
We probably wouldn't have brought it back at all.
...
In December 1990, the research institute held its annual achievement reporting meeting.
I organized the data on F-47 into a separate report and handed it to Professor Kijima.
The professor read it very carefully.
He admitted that some traits of F-47 were indeed of high research value.
But as a forage grass, its problems were also quite obvious.
Insufficient protein.
Poor lodging resistance.
Inability to set seeds normally.
High reproduction cost.
Especially the last point.
A forage grass that required artificial division to reproduce would be extremely troublesome to popularize, no matter how good its quality was.
Therefore, the professor ultimately decided to include F-47 in the special germplasm preservation list and withdraw it from conventional forage candidate evaluation.
I agreed.
Because from the perspective of forage research, his judgment had no problems.
Our project was to improve cattle feed for S-Farm.
Since there were other candidate varieties with more balanced performance, limited research funds should be spent on those varieties.
Thus, F-47 was preserved.
I originally thought its story ended there.
Until the winter of the following year.
...
January 17, 1991.
That morning, the TV in the research institute's cafeteria was broadcasting news of the U.S. airstrikes on Baghdad.
I sat in the corner eating breakfast, looking at those lights in the night sky on TV, and didn't pay too much attention to it.
After all, the place where the war was happening was simply too far from Hokkaido.
But not long after, the logistics department notified us that the kerosene supply for the greenhouse had gone up in price.
The head of the office held the new procurement quotation and complained in the hallway for a good long while.
Only then did I realize that the war happening in the Middle East would ultimately affect the heating cost of a small greenhouse in Shakotan.
However, this matter didn't make me think of any new energy sources at the time.
I studied crop breeding.
When oil prices rise, of course the people responsible for energy should figure out a way.
What truly changed my mind was an ordinary digestibility determination a few weeks later.
That night, I stayed behind in the laboratory to work overtime.
There were still many materials to be measured.
F-12, F-88, F-103, and several control varieties prepared for elimination.
The procedure was pretty much the same.
Grind the plant samples, process them first, then add cellulase, place them under constant temperature conditions, and finally measure the remaining undissolved matter.
I had done this kind of thing many times.
People who study forage simply want to know how much stuff can be utilized by animals.
But that night, looking at a row of samples on the desk ready for residue determination, I suddenly thought of a question I hadn't seriously considered before.
We had always been looking at how much was left.
Then, where did those decomposed plant components actually go?
A considerable portion of them should have already been converted into soluble sugars and other substances.
That being the case, why not directly measure the sugar in the solution?
This idea wasn't really anything extraordinary.
The fact that plant cellulose can release sugars through enzyme treatment was something people had studied long ago.
But for someone who only cared all day long whether cows could digest forage, looking at an experiment he had done hundreds of times from a different perspective still felt somewhat magical.
I almost immediately thought of F-47.
Its digestibility was 61.3%, after all.
Ordinary Miscanthus was only around 39.8%.
If these differences could be converted into more available sugars, plus the already high soluble sugar content in its stems...
I decided to give it a try.
...
On February 19, I took out the dried F-47 samples preserved last autumn.
The control was still the ordinary Miscanthus genus material F-12.
Using the institute's existing enzymes and equipment, I additionally conducted a set of exploratory saccharification tests.
Forty-eight hours later, the results were out.
Fermentable sugars released per 100 grams of dry matter:
F-47: 33.8 grams.
F-12: 17.2 grams.
Nearly double.
I stared at these two numbers for a long time.
Because prior to this, I already knew that F-47 had a very high digestibility.
So appearance of such a result didn't seem very strange.
But when the result was truly laid out before me, I was still somewhat excited.
In March, I managed to get some brewing yeast.
It was obtained through a college classmate from an acquaintance at a winery in Yoichi.
The other person asked me why someone who worked with forage needed yeast.
I said I was preparing to do feed experiments.
This answer wasn't entirely a lie either.
After all, I really was researching feed in the beginning.
What I did later didn't have much to do with cows anymore.
I inoculated the F-47 saccharification liquid with yeast and fermented it at thirty degrees.
Seventy-two hours later, the glucose was basically used up.
However, quite a lot of xylose was left over.
Ordinary brewing yeast had no way to effectively utilize xylose, which was not a new problem at the time.
I didn't solve it either.
But ethanol was indeed produced.
Converted to dry matter, about 11.9 grams of ethanol could be obtained per 100 grams.
Using the laboratory's small distillation apparatus, I produced about twenty milliliters of crude alcohol.
Twenty milliliters.
To be honest, pathetically little.
Not even enough to fill an ordinary wine glass.
Yet I held that small bottle and sat in the laboratory for a long time.
Because I suddenly realized that for over a year, I had been evaluating F-47 using a standard that might not be suitable.
I wanted it to become high-quality cattle feed.
But who stipulated that a plant must become feed?
From then on, I began to seriously consider—
whether F-47 could be used as a plant raw material exclusively for fuel production.