Author Archives: Ulrich Riemann

Interspecific BC1 Jatropha hybrid

The Bionic Jatropha Breeding Platform in 2013

We have already passed the first half of February of this year, so this article is overdue, if we want to make this kind of posting a tradition. See last years version here: The Bionic Jatropha Breeding Program in 2012.

First of all let’s summarize some 2012 highlights one last time:

Interspecific BC1 Jatropha hybrid

Interspecific BC1 Jatropha hybrid

In December we have reached the BC2 stage following our interspecific breeding path. The first batch of BC2s are all descendants of an early and white flowering BC1, the female parent of which is in turn our non-toxic, male-sterile Jatropha plant from the wild in Guatemala. The pollen donor was an interspecific hybrid crossed from an elite toxic JcL cultivar and Jatropha integerrima. Many more genetically diverse BC2 hybrids will follow over the course of this year as our large population of different BC1s gradually start flowering. Some actually have done so by now.

Quite unexpectedly earlier in 2012 we observed enormous heterosis effects (hybrid vigor) in several different intraspecific hybrids we had kind of neglected. Some increasing seed weight compared to their parents by up to 100%. A finding which made us gravely rethink and adjust our breeding processes and objectives. Later we then saw heterosis again in another hybrid with fruit clusters consistently around 50 fruits and seed weight still significantly above the parental average. All three of those hybrids initiated flowering only 6 months after seed germination. These plants actually convinced us that consistent seed yields of 10 kg and more are within reach. We do expect to see a lot more of positive heterosis results as we only started to fully exploit our germplasm collection by crossing our genetically most distant accessions in the late months of 2012.

Two more areas are worth mentioning where we expanded our knowledge of Jatropha development significantly during 2012, possibly taking us to a global leadership position in Jatropha R&D: Grafting and multi species (more than 2) Jatropha hybridization. We have made great progress on both work streams and do expect major break-throughs to be reported by the end of this year.

BC1: 7.10xH1.1

Unusual Jatropha flowers on a hybrid BC1

The walk through of our 2012 results actually already covers most of the outlook for 2013: our interspecific hybrids will most likely reach BC4 stage moving closer to possible commercial exploitation through selection from a widening population and dedicated heterosis breeding will teach us all we need to know to develop a first line of F1 hybrids in 2014 from our germplasm range.

The first 6 weeks of 2013 gave us an extra boost in confidence looking at the great developments we saw in the usually not very eventful core dry season here in Ghana. This is symbolized by the beautiful flowers on the left, another BC1 which set flowers for the first time in January which were manually pollinated and emerged into 4 fast growing fruits by now.

SGB Jatropha patent application published

A somewhat confusing patent application by SG Biofuels Ltd (KY), a Grand Cayman Islands company, has been published recently in several versions. Here is the official European link:

EP2521438  (A1) – 2012-11-14 - JATROPHA HYBRIDS THROUGH FEMALE ONLY TRAIT

And the US Version: US2012324783  (A1) – 2012-12-27

Several further international applications exist and a provisional US application No. 61/292751, dated January 6, 2010 is quoted for priority rights.

Explaining all the weird technicalities of international plant patents and the differences between US and European rules is not the purpose of this article. However, it should be noted, that the claims between the European and the US version differ substantially, obviously in order to adjust them to the corresponding requirements and take maximum advantage of them. As priority based on the US provisional application is claimed also for the European version the differing claims might become a serious technical hurdle for the issuance of a European patent.

On a side note, if you want to know more about patenting plants in Europe have a look at this excellent presentation “Latest Developments in Patenting Plant Inventions in Europe”.

Putting law and regulations aside, lets analyze the claims a little bit:

  1. “A Jatropha curcas plant characterized by an inflorescence with only female flowers.”  is the first central claim. Later on they call this the “FO trait” in an obvious attempt to avoid the common term “male sterile”. So in essence, this seems to be a business idea, to claim IP rights to all male sterile Jatropha plants and their use! It is absolutely possible within US and European rules to patent a certain plant trait. Now lets see, an invention needs to be “novel and inventive teaching”. That’s why they claim, their “FO trait” somehow emerged during their breeding activities. That would definitely make it “novel and inventive”. However, later they also claim the plant with the “FO trait” to be a homozygous (relative to the trait) and an inbred line. That is technically impossible for a male sterile, at least if it derives from an “novel and inventive” cross.
  2. Further, a wild plant can never be patented, even if it is discovered for the first time. That is also true for all traits of said wild plant. As we want to keep ourselves and fellow Jatropha breeders safe for the future, we have no choice but to disclose: Bionic Palm is in the possession of several male sterile Jatropha plants deriving directly from seeds that have been collected from wild plants at the center of origin. Specifically one of those plants has been grown from seeds collected from a wild Jatropha curcas L. bush in the South-West of Guatemala. We are in the possession of exact GPS coordinates of the mother plants location. The behavior of that male sterile plant is completely identical with anything described as the “FO trait” in the SGB patent application. (In the next few days we will publish a document which will fully disclose all our findings regarding our male sterile accession from Guatemala origin)
  3. Further, the SGB patent claims ownership over a most common and widely taught production technology for F1 hybrid seeds: a planting pattern for a seed farm combining a male sterile accession with another inbred superior accession for natural pollination. The only part “inventive and novel” in this claim is, that such a technology is combined with the word “Jatropha”. Other than that this could be copied straight from a breeders handbook! This seems to be an attempt to force major parts of a future Jatropha industry into paying royalties to SGB in case the application would be successful.
  4. And finally it is very interesting to see the new direction the claims in the US version are taking. Here the patent application seeks control over all derived fuels from Jatropha seeds originally produced using the above method. That is driven by the idea, that where ever in the world the seeds have been grown, once the product enters the US and the seed production method applies royalties can be forced on the importer.  

What can be the purpose of an otherwise meaningless patent like this one?

I want to leave the answer to this question to the reader and just ask 2 simple questions:

  1. Is the ailing Jatropha industry ready for patent wars as we see them unfolding in the mobile handset or the software industries?
  2. Is SGB trying to follow a Monsanto principle? Roundup and Canola are still well known at least to the agric experts…

What do we at Bionic Palm intend to do?

Well, we will do what ever we can to keep these applications to be turned into patents. We believe, publicly disclosing the ownership of above mentioned wild male sterile from Guatemala should do the trick. But we will also make sure, that this disclosure document will get to all patent offices concerned.

We will definitely keep you posted on any further developments.

 

Jatropha, a plant with a future – new report from Hardman&Co

Its report season… another one on Jatropha has just been published. Actually this one I had awaited for several months. Finally it has been released today. They titled it “Jatropha – Plant with a future” and it has been released by Hardman&Co, a UK based corporate research specialist for the financial industry. The PDF can be downloaded here or directly at their website.

After their excellent last report on Jatropha investment expectations were running high for this latest edition focusing exclusively on the development of improved Jatropha planting material. However, I find myself a little disappointed as this report mainly focuses on a handful of well known developers and obviously their response to telephone interviews. While the information contained is definitely interesting and well researched, I miss a more independent assessment of the actual development status and the possible paths forward for the evolving industry.

My disappointment was even greater when I realized that the huge global scene of independent developers have not been mentioned with one word. I have been watching the handful “incumbent” organisations mentioned in the report over the years and I don’t believe any of them will deliver the break trough developments the industry is desperately waiting for. Instead, I expect a lot more from the loosely connected modern networks which are working on Jatropha around the globe. What I see is somewhat similar to modern open-source software development. We are an active member of this scene.

Admittedly it is more difficult to explore that parallel universe of Jatropha development, because proper research requires a lot more insight into the subject itself. But it is also the only way to figure out and report what’s really going on in Jatropha development  Therefore I am seriously contemplating about sitting down for a couple of days myself to complement this report with a second one from the independent networked Jatropha development domain.

Actually, it’s the main purpose of this very website too to share more information about Jatropha development when it becomes available. So after reading the report please come back here and let’s discuss some of the “facts”:

For example, the world market price of soybean meal is not USD 300 (as claimed in the report) but actually over USD 450 per metric ton.

Or why is it, that we have announced, improved, high yielding planting material can be made available to any project within 18-24 months via our localized client site breeding services? The report says it will take to the end of the decade for improved material to become widely available! An unacceptable timeline for anyone interested in Jatropha farming today.

Heterosis seed comparison

Jatropha hybrid heterosis: 50 fruits in a single cluster

Jatropha hybrid heterosis

50 fruits in one cluster

We are currently observing another impressive result of hybrid vigor in our Jatropha Breeding Program: a six months old hybrid exhibits the first fruit cluster with exactly 50 healthy fruits. The seedling germinated on March 17 (my birthday :-)) and has reached a height of more than 2.5 meters by now. Manual pollination has been conducted 10 days ago.

Among other this hybrid also shows a distinctively superior tolerance to drought stress than both parents. As usual drought tolerance goes together with relative (horizontal) pest resistance.

2 sister plants of the identical hybrid which had germinated in the same week are showing similar impressive fruit counts for their first clusters: 35 and 42 respectively.

CLO7.10xGHLT hybrid

Hybrid fruit cluster

Update: the same fruit bunch 10 days later…

 

 

 

 

 

Update 2: On November 23 we harvested the first fruits from this bunch. Obviously, we were very curious to find out about a possible heterosis effect on seed size and weight. Well, what can I say? It’s another breakthrough really that would deserve its own separate article on this site:

We weighed 15 freshly harvested seeds at a total of 18 grams. This converts into a 100-seed weight of 120g. That’s a new record in our breeding program.

Heterosis seed comparison

Seed comparison with parental accessions

As a matter of completion, the 100-seed weights of the parents:

  • female parent (top left): 90g
  • male parent (top right): 45g.

It will be interesting what we are going to see in the next generations of hybrid crosses. The plant has already been crossed with most of our high quality accessions and also back crossing with both parents has begun. We will definitely report again in the future. 

Jatropha curcas scion grafted on Jatropha gossypifolia rootstock

Jatropha curcas graft with Jatropha gossypifolia rootstock

We wrote about the role of grafting in Jatropha development before. Today we return to a grafted plant at our test farm. It is almost 3 years old and standing strong. When we created it a random scion was cut from one of our standard field plants and grafted on a cut Jatropha gossypifolia with kind of a reverse cleft graft.

A main property of this grafted plant is an immensely increased drought tolerance compared to Jatropha curcas samples in the vicinity. The stem appears slightly thinner (it’s on the left side in the picture. The main stick in the center is a support because the plant is exposed to constant strong winds), but is developing.

The graft is now constantly flowering and fruiting and develops a unique canopy structure. It demonstrates the many yet unexplored options available for Jatropha agronomic methods. Applying advanced grafting techniques as they are used in modern fruit tree development is only one of them. This graft was one of our first three tests ever conducted. An experienced grafting expert could have definitely done a much better looking job…

We have added another interesting picture which indicates clearly the different textures of the stem below and above the graft. More expertly done the graft would probably be closer to the ground.

Stem of grafted Jatropha

Stem of a grafted Jatropha 30 months old

It should be noted that this plant suffered from heavy mealy bug attacks lasting for almost 18 months. Development basically came to a halt during this period. The plant exhibited above average strength in surviving the attack and was one of the first in the area which recovered and started flowering and fruiting again.

The earlier article discussed in detail the potential of grafting for accelerated farm development and recovery of suspended Jatropha projects with future elite cultivars. At Bionic Palm we see great potential in such an approach and keep experimenting with it while we move along with our hybrid breeding program.

 

 

 

 

 

Heterosis effect showing in seed size

Heterosis increases Jatropha seed weight 100% and more

It has been only a month since we last wrote about heterosis in Jatropha breeding. However, we had not seen then, what we have seen now…

We have harvested a first batch of Jatropha seeds from the intraspecific hybrid exhibiting by far the strongest heterosis effects in leave size and height. And we were up for a real surprise.

We had recently seen 2 scientific reports on heterosis trials with Jatropha in South East Asia describing maximum positive heterosis in seed weight of up to 25%. We therefore never expected to see our highest 100-seed weight increasing so much. Extrapolated from 20 harvested seeds it slightly exceeds 110g. Size can be compared well in the picture above with control being a typical average size seed at a 100-seed weight of 56g.

Our own findings are completely in line with the above mentioned reports, while showing even further potential. We still have to cross those parental lines in our possession with the greatest distance according to our genetic map. As all available results point to the very strong positive correlation between heterozygosity and heterosis in intraspecific Jatropha hybrids we expect that our latest findings can easily be exceeded once all our planned hybrid crosses have fruited.

This latest success supports our ongoing planning for a dedicated heterosis breeding sub-program which could allow for stable, highly superior F1 hybrids within less than 2 years.

 

Large diversity in F2 interspecific hybrids not unexpected

Jatropha interspecific hybrids F2 generation available

Fruit on an interspecific hybrid

Fruit on an interspecific F1 hybrid

In the second half of August we successfully germinated the first three F2 generation seedlings from one of our interspecific hybrids. From now on we have fast growing F2 Jatropha interspecific hybrids for further breeding. While some research reports indicate that this type of interspecific hybrid is sterile or not interesting for breeding, we observe ongoing successful seed development on several of our early F1 plants deriving from different Jatropha curcas female parents. So we expect more additions to this F2 generation population in the near future.

As expected from applying simple Mendelian rules the F2 generation phenotypically exhibits much more diversity than the F1 generation.

All of the 3 seedlings we have so far appear strong and healthy. However, only the one on the left hand side seems to be as fast developing as expected while the other 2 are only moving along slowly.

Large diversity in F2 interspecific hybrids not unexpected

F2 seedlings of interspecific JcL hybrids 1 month old showing wide variation

We expect our F2 interspecific population to grow very fast over the coming months involving many different F1 interspecific parents with genetically very different Jatropha curcas parents in turn. Currently many F1s are initiating their first flowering. On the interspecific route of our breeding program this is really what we have worked on for almost 3 years. We have clearly begun moving into terrain which has not previously discussed in the scientific literature to the best of our knowledge.

This development marks another success in achieving our initial target to bring as much variability into the germplasm available to our Jatropha breeding platform for future improvements through selection. We are now able to start work on a real ground breaking next step we had been planning for since we did the first interspecific cross almost 2 years ago. With a little luck first results will become visible before the end of the year.

Further observations on the interspecific breeding route will be published here as much as possible, but of course we will have to start protecting our intellectual property where we reach the forefront of commercial hybrid development as we are expecting a meaningful patent application before too long.

Jatropha seed comparison

What makes an elite Jatropha cultivar

What exactly makes a certain Jatropha accession an elite cultivar? You can find the term everywhere but a clear definition is usually not given. Therefore I want to explore this subject today and attempt to demonstrate how an answer translates into  the daily work of a Jatropha breeder.

The term itself is simple: elite means something like “better than average” and cultivar is just an abbreviation of “cultivated variety”. But what does it really mean to us?

The most general and holistic objective when breeding for an improved commercial crop is of course always maximum profitability for the farmer. As this is self explanatory only a next level break down of objectives can offer some insight. In our program we use 4 groups of traits which in turn again break down into the real, identifiable traits when observing a selected plant.

The groups we use at Bionic Palm are:

  1. Seed Quality
  2. Seed Yield
  3. Agronomy
  4. Pest resistance.

We are looking for improved traits in all of the above groups. Let’s have a quick look at each group to see some of the aspects a breeder is really looking for every day.

Seed Quality
For us this includes several individual traits, all linking back to profitability of the crop of course.

  • Toxicity
  • Uniformity within the seeds
  • General healthiness of the seeds
  • Chemical composition of oil and cake.

No further explanations needed here I think.

Seed Yield
This is the most relevant for most people discussing Jatropha, but what are we really talking about, when we use the term? 

Many people would just want to count the seeds or go by tons per ha. An approach that would translate into something like number of seeds harvested per plant per year. This again translates into bottom line traits like branching (self or pruning induced), number of fruit clusters or fluorescences per branch, number of fruits per cluster (= number of total flowers in a fluorescence plus male:female flower ratio). Not too difficult to understand, once you know the Jatropha exclusively flowers at the top of a branch.

However, that’s not even half the truth. In our understanding the high level measurement must actually be average amount of crude oil obtained per ha (or acre) combined with average amount of cake per ha.

We have recently taken a closer look at seeds obtained from the various hybrids we produce. A comparison shows significant differences in seed weight and oil content which are actually quite stunning. And the spread is increasing the longer we breed…

Jatropha seed comparison

The typical measurement 100 seed weight varies from around 45g to 98g in our current accessions and oil content ranges from 25% to clearly over 50%. That means with an equal number of seeds per ha I can actually have up to 4 times the oil!

Agronomy

 Lets move on to traits important for agronomic reasons. Many of them are related to the cost involved in farming Jatropha and the most to the ease of harvesting.

There is the uniformity of fruit ripening which determines how many times a picker has to pass a plant per season, then there is canopy structure and size of fruit clusters which determine the ease and speed of harvesting. Other important agronomic traits are self branching (without the need of pruning). Also drought resistance or rather the reaction to water stress is an important trait. We find that there are tremendous differences between accessions. The difference is at what level of water stress a plant stops female flowers, no flowers at all, throws off developing fruits and more. We see a clear pattern of those reactions differing greatly among accessions. I also have to admit that so far the best performers are the weakest in terms of water stress resistance.

Pest Resistance

Many of the common publications about Jatropha suggest that their toxicity would protect them from pests. This is not the case! The most serious pest we are confronted with is the papaya mealy bug. It comes and goes with water stress. There is no correlation with toxicity at all, however a clear correlation with resistance against water stress. And it should be at least mentioned here that the available phosphorus in the soil plays a major role in any plants ability to handle water stress. Therefore the right soil amendments like mycorrhiza can make a huge difference in addition to the best suited accessions if you plan to plant Jatropha in the desert. :-)

I am aware that this review of relevant traits in Jatropha is anything but exhaustive, but I don’t want to bore my readers too much with the obvious. Therefore I tried to point out only a few of the less discussed but important aspects of this fundamental subject on the way to true Jatropha elite cultivars.

The role of heterosis in Jatropha breeding

Today I want to take a closer look at the relevance of heterosis in Jatropha breeding. Heterosis or hybrid vigor, or outbreeding enhancement is “the increase in growth, size, fecundity, function, yield, or other characters in hybrids over those of the parents”. Find more information on heterosis on Wikipedia.

Heterosis effect can be forecasted based on this genetic variability map

Almost 3 years ago we started all our breeding efforts from this genetic variability map created by our friend and advisor Tommaso Barbi working at Geneticlab in Trieste, Italy at the time.

The parental accession base of the Bionic Palm Jatropha breeding program is not a 100% overlap, but we do have quite a significant number of key accessions in our program which are also shown in the map while many others can be linked to one of the clusters genetically or phenotypically.

Simply speaking, crossing more distant accessions from the map increases the likelyhood of significant variability in the hybrids genetically expressing itself in a higher degree of heterozygosity. Another crazy insider term from molecular biology, I know. Heterozygosity describes the amount of differences between corresponding alleles in a DNA sequence. Homozygosity is the term for the opposite situation of corresponding alleles being similar.

It has been found that heterozygosity usually correlates with strong heterosis effects. In the map above a cross between more distant accessions leads to a higher heterozygosity in the resulting hybrids and therefore higher levels of heterosis can be expected. Only about 6 months ago we started to be able to observe different heterosis levels in our hybrids.

The molecular basis of heterosis is still not fully understood. Current believe among scientists is that healthy alleles across numerous genes from one parent compensate for dysfunctional alleles at those genes from the other, distantly related parent (and vice versa for a second set of genes). A plant exhibiting a high level of heterosis would thus be the genetically most healthy.

This explanation fully corresponds with the opposite effect of inbreeding depression. Here parts of the parental alleles become dysfunctional or silenced because there is too little genetic difference between the two parents. The very strong expression of heterosis we observe when crossing a semi-commercial Jatropha accession from Africa or Asia with a wild accession from the center of origin therefore suggests at least a mild inbreeding depression on a very narrow genetic variability in the germplasm originally introduced by the Portuguese seafarers when they first distributed Jatropha around the world.

So far observations are limited to leave size and, at a smaller extent, stem diameter. Over the coming month fruit and seed sizes will become visible too. With the leaves we quite commonly measure lengths of up to 20cm where the usual average is no more than 10cm.

The following picture demonstrates the phenomenon very clearly.

Heterosis in Jatropha breeding can be observed in this intraspecific hybrid plant

The small plant in the front germinated in November 2011 and is a cross of 2 accessions with distinct phenotypic differences but, genetically very similar, they are both from the upper left cluster in the map. The much larger plant behind in the center germinated in March 2012, 5 months later, and represents a cross between the upper and lower left hand clusters. Average leave length is almost double that of the plant in the front and of course the overall size is significantly different.

 A heterosis effect is also visible in many of our interspecific hybrids, but notably less so in those which have a very short seed-to-seed time of less than six months. A behavior which can be attributed to negative heterosis (=shorter seed-to-seed cycle). Interspecific heterosis can be observed in the following picture.

Heterosis in Jatropha breeding

The plant on the far left hand side shows by far the most dramatic heterosis effects. Followed by the next 2 plants to the right with heterosis also clearly noticeable. All 3 happen to be non-toxic interspecific hybrids. Then we see an early flowering and fruiting interspecific hybrid exhibiting no visible heterosis. Similarly at last, on the far right some regular Jatropha can be seen. From left to right we go from high level heterosis to none at all.

In front a rare chili pepper can be seen which is currently tested for drought resistance.

The findings described in this post have been highly unexpected. We are currently waiting for the first seeds from plants exhibiting strong heterosis to mature so we can check seed weight and oil content. Based on the results we will have to decide if we are going to open up a new heterosis breeding route in addition to our current approaches.

Early BC1 collection

Jatropha hybrid backcrosses available for further breeding

Before our proof-of-concept phase officially came to an end on June 30th, 2012 we succeeded in taking our efforts to the next level. Our first Jatropha hybrid backcrosses have germinated and reached seedling stage by mid June. This achievement marks an enormously important step forward for the Bionic Palm Jatropha breeding program. 

Jatropha hybrid backcrosses exhibit enormous genetic variability even at the seedling stage

The above picture shows the huge variability among our earliest BC1 seedlings. Over the coming months this collection will grow into the hundreds. We aim for a BC1 population incorporating all of our original parental germplasm and most of its cross combinations. Thus we will gain the maximum possible genetic variability base which we can later draw from when we breed for truly superior commercially usable hybrids.

What we define as our BC1 population is the result of back-crossing interspecific Jatropha curcas x Jatropha integerima hybrids into our parental Jatropha curcas accessions.  This is a standard breeder’s approach if he wants to transfer specific traits (certain properties) from a related species into his species of interest. However, we are not into pedigree breeding at this time. Our primary aim is still to increase genetic variability of our population to the utmost possible.

We are looking for the greatest possible heterozygosity in these plants which can be assumed to be 100% in many cases. According to scientific research reports on the genetics of Jatropha this can only be achieved through an interspecific breeding route. While this kind of approach is very time consuming we do expect to gain a substantial shift in the Jatropha genetics.