Arxiu d'etiquetes: hybridization

How is genetic engineering done in plants?

For years, by crossing, scientists have achieved plants with a desired characteristic after many generations. Biotechnology accelerates this process and allows to catch only the desired genes from a plant, achieving the expected results in only one generation. Genetic engineering allows us to do all this. In this article I will explain what it is and how does it work.

WHAT IS GENETIC ENGINEERING?

Genetic engineering is a branch of biotechnology that consists in modifying hereditary characteristics of an organism by altering its genetic material. Usually it is used to get that certain microorganisms, such as bacteria or viruses, increase the synthesis of compounds, form new compounds or adapt to different environment.

It is a safer and more efficient tool for improving species than traditional methods (crossing) as it eliminates much of the randomness. On the other hand, modern biotechnology also becomes a new technology that has the power to modify the attributes of living organisms by introducing genetic material prepared in vitro.

It could be defined as the set of methodologies to transfer genes from one organism to another and express them (to produce proteins for which these genes encode) in different organisms of the original organism. DNA which combines fragments of different organisms is called recombinant DNA. Consequently, genetic engineering’s techniques are called recombinant DNA techniques.

Currently there are more plant organisms genetically modified than animal organisms. For this reason I will explain genetic engineering based on plants.

GENETIC ENGINEERING vs. TRADITIONAL METHODS

This methodology has 3 key advantages compared with traditional methods of genetic improvement based on hybridization:

  • The genes could come from any specie (for example a bacteria’s gene can be incorporated in soy‘s genome).
  • At genetically improved plant you may introduce a single new gene preserving the remaining genes from the original plant to their offspring.
  • This modification process delays less the deadlines than improvement by crossbreeding.

With this way you can modify properties of plants more broadly, more accurate and faster.

In traditional crossing it generates a hybrid which combines randomly genes of both parental organisms, including the gene of interest encoding the desired trait. In contrast, biotechnology techniques only pass one or few genes which encode a specific trait known. The new plant has all the original genes of the plant and an introduced gene accurately and directed (Figure 1).

fig1ENG
Figure 1. (A) Traditional method where, by crossing, a new variety is obtained. This carries the gene of interest (red), but also another genes randomly. (B) With genetic engineering we obtain a new variety of commercial plant with the gene of interest (red) of any other species (Source: Mireia Ramos, All You Need is Biology)

METHODOLOGY OF GENETIC ENGINEERING

Obtaining a transgenic organism through genetic engineering techniques involves the participation of an organism who gives the gene of interest and a receptor organism who will express the desired quality. The steps and the process techniques are:

0/ DECIDE THE AIM: MAKE KNOCK IN OR KNOCK OUT

KNOCK OUT:

This technique is to remove the expression of a gene, replacing it with a mutated version of itself, this being a non-functional copy. It allows the gene is not expressed.

KNOCK IN:

It is the opposite of the knock out process. A gene is replaced by a modified version of itself, which produces a variation in the resulting function of it.

In medicine, the knock in technique has been used as a strategy to replace or mutate genes that cause diseases such as Huntington’s chorea, in order to create a successful therapy.

1/ DOUBLE CHECK THAT THERE IS A GENE CODING FOR THE CHARACTERISTIC OF INTEREST

Firstly, you have to check the characteristic of interest comes from a gene, as this will be easier to transfer to a living organism that does not.

2/ CLONING THE GENE OF INTEREST

It is a complex process, but outline the steps are the following:

  • Extract DNA
  • Find a gene among the genes of this DNA
  • Sequence it
  • Build the recombinant vector

The DNA of interest is inserted into a plasmid, a circular DNA molecule with autonomous replication. The plasmids of bacterial origin are the most used (Video 1).

Video 1. “Clonación de un gen en un plásmido vector”. Explaining the use of plasmids as a vector in the process of cloning (Source: YouTube)

The development of these techniques was possible by the discovery of restriction enzymes. These enzymes recognize specific sequences and cut the DNA by these points. The generated ends can be sealed with ligase enzyme and to obtain a new DNA molecule, it called recombinant DNA (Figure 2).

adnrecombi
Figure 2. (1) Plasmid’s DNA (2) DNA from another living organism (3a, 3b) The restriction enzyme cuts DNA (4) The restriction enzyme recognizes AATT sequence and cuts between A and T nucleotides (5) The two DNAs are contacted with the purpose of forming recombinant molecules (6) A ligase enzyme joins the DNA ends (Source: GeoPaloma)

3/ CHARACTERISE GENE OF INTEREST

If we know the gene sequence we can compare this sequence with known gene sequence through bioinformatics, provided to determine which gene looks and assign a possible function. So when we have predicted the function of cloned gene we confirm it in vivo, usually transferring it to a model organism.

4/ MODIFY GENE OF INTEREST

We can add (promoter, introns…) or mutate sequences inside the encoding region.

5/ TRANSFORMATION OF A LIVING ORGANISM WITH GENE OF INTEREST

When we have finished the gene building with the desired gene and the promoter, the recombinant DNA is inserted into the cells of the living organism that we want to modify.

6/ CHARACTERIZATION GMO

When we already have the GMO (Genetically Modified Organism) it is analysed from the molecular and biological point. In the molecular analysis it must demonstrate if you have one or more copies of the transgene or how and what tissues the gene is expressed. In the biological analysis it looks if it achieves the objective for which it was designed.

REFERENCES

MireiaRamos-angles

Ancestors they didn’t teach you in school

Surely you know any of the following names because they are classic ancestors we learned in school: Lucy, Homo habilis, Homo erectus, Neanderthals… but our history has many more players, and every so often new discoveries are made that change our lineage tree. Find out in this article the latest findings your teachers could not explain to you .

HOMO NALEDI

Reconstrucción facial de Homo naledi por John Gurche. Foto de Mark Thiessen.
Homo naledi’s facial reconstruction by John Gurche. Photo: Mark Thiessen.

It is almost forced to start with one of the latest discoveries that is encouraging discussions in paleoanthropology to gain a key position in our family tree. The discovery of a new species, Homo naledi, was published the September 10, 2015 by Lee Berger et al. It was discovered in a cave system in South Africa named Rising Star at the Dinaledi chamber (“Naledi” means “star” in the local language, Sesotho). It is especially interesting for several reasons:

  • At the moment in the site have been found more than 1,700 human fossils accumulated, making it the largest of South Africa, behind the famous Sima de los Huesos (“Pit of Bones”, Atapuerca, Spain), the largest of the world, with more than 6,000 fossils.
  • The cave is very difficult to access, with corridors of only 19 cm wide, so it was a selected team of 6 thin paleoantropologysts (all women) that reached them.

Esquema del sistema de cuevas de la cámara Dinaledi. Imagen de Jason Treat, NGM Staff, NGM maps, fuente: Lee Berger, Wits. Tomada de National Geographic.
Scheme of the cave system of Dinaledi’s chamber. Image by  Jason Treat, NGM Staff, NGM maps, Source: Lee Berger, Wits. Adapted from National Geographic.

  • The bones belonged to 15 individuals of all ages, male and female, so we can get extensive information about the new species. Some were even on the floor of the cave without mineralize.
  • The physical characteristics of H. naledi are a mix of Homo traits (height, feet) and Australopithecus (shoulders, chest, pelvis), the genus from which most scientists believe Homo appears about 2.8 to 2, 5 million years ago. This may suggest that H. naledi could be the first Homo, the missing link between Australopithecus and us.

    Una parte de la impresionante cantidad de huesos de Homo naledi descubiertos. Foto de John Hawks
    Some of the impressive number of bones discovered Homo naledi. Photo by John Hawks
  • The most intriguing of this discovery, it is believed that the bones were placed there deliberately. By geography, access to the cave was the same as today, they could not fall into the pit, the bones could not be brought by a water flooding or any animal, they have no marks of violence … It could be a funeral ritual? So far, the first rites are attributed to H. neanderthalensis, with most modern physical characteristics and a large cranial capacity compared to H. naledi (1.475 cm3 versus 560 cm3  at the most).

The oldest known Homo fossil, 2.8 million years old, corresponds to a jaw found in Afar in March 2015 which has not been associated to any species. Was H. naledi the first Homo? Is it really an ancient species? Is it possible they had self-awareness so early and cared for their dead? Unfortunately, researchers have not been able to date the remains yet, so many questions remain unanswered and we will must wait for future interpretations of one of the most important discoveries of recent times.

THE DENISOVANS

In Denisova Cave (Siberia) in 2008 was found a non-spectacular fossil: a piece of a finger bone that was dated 30,000 years old and attributed to an individual of about 8 years which turned out to be a gir. But when DNA was extracted, it was concluded that belonged neither to H. sapiens or H. neanderthalensis, but to a new species. Later two teeth of another individual of the same population were found. In the same cave also Sapiens and Neanderthal remains were found.

Diente, muela, denisova, denisovanos, teeth, tooth, denisova
The denisovan teeth.  Photo by Max Planck Institute.

Is it possible that Denisovans hybridized with Sapiens? DNA studies in the current populations indicate that 5% of DNA aboriginal Australians, Papuans and other peoples of Melanesia is Denisovan. On the other hand we know that 20% of the DNA of accumulated European populations is Neanderthal.

WHERE DO WE LOCATE THEM IN OUR FAMILY TREE?

It is thought that Neanderthals and Denisovans had a common ancestor (H. heidelbergensis), who emigrated to western Europe and Central Asia evolving to H. neanderthalensis, who subsequently hybridized with us, and from Southeast Asia where would evolve in the hominin Denisova, who also hybridized with H. sapiens. This would explain the presence of DNA in the current populations of Australasia.

HOW THEY WERE LIKE?

The absence of more fossils or traces of objects and tools prevent us to know how they looked like and what were their skills. Nor it has been found explanation for the lack of Denisovan DNA in the Russian or Chinese populations, so close geographically to the Denisova cave. Denisovans remain a mystery to science.

THE FLORES WOMAN

Homo floresiensis. Reconstrucción de John Gurche
Homo floresiensis. Reconstruction by John Gurche. Photo by Chip Clark

Homo floresiensis, as its name indicates, lived on the island of Flores (Indonesia) only between 95,000 and 12,000 years ago. It was discovered 12 years ago. It is the only site where this species is found.

As in previous fossils, the mix of features caught the attention of the scientific community, especially for its small cranial capacity and height, earning them the nickname hobbit. First they thought it was an individual with a pathology, or a pygmy of a known species, as their morphology was very strange in a so modern fossil. But now we have remains of at least 12 individuals with the same traits, so we can talk (for the moment) of another species.

HOW THEY WERE LIKE?

  • Small height: the most complete skeleton belongs to a female only one meter tall and 25 kg weight.
  • Small skull: their cranial capacity (380-420 cm3) was similar to the current Australopithecus or a current chimpanzee, but the brain had a more similar Homo anatomy. The teeth were large relative to the skull.

Reproducción de cráneo de Homo floresiensis. American Museum of National History. Foto de Mireia Querol
homo floresiensis (LB1) skull cast. American Museum of National History. Photo by Mireia Querol

  • Long feet and short legs: feet were very long in relation to the legs, which were short and stout. This and more features suggest that locomotion was different from ours and were bad runners.
  • Long arms: besides a proportion nearest to Australopithecus and H. habilis than H. sapiens, arms were robust and had a powerful musculature.
  • Stone tools and fire: besides the existence of tools of earlier hominans found in the cave, some tools have been associated to H. floresiensis with a technology similar to the Oldowan Industry, the first to be invented. Also they dominated the fire.

WHY THEY WERE SO SMALL?

Controversy continues: was a direct descendant of Australopithecus (how could they have traveled so far from Africa?), or a recent member of our family tree so small due to lack of resources?

The insular dwarfism is an evolutionary process due to a long-term isolation in a small area with limited resources and lack of predators. Flores pygmy elephants (Stegodon) hunted by H. floresiensis  with this adaptation were also found. The opposite process it is the island gigantism, in which animals that are usually small on the continent are giants in the islands, such as the Galapagos turtles and the extinct lizards or rats of Flores.

Un lagarto gigante se enfrenta al hombre de Flores. Imagen de National Geographic
A giant lizard faces Flores man who has caught a rat. Image by National Geographic

H. floresiensis may be the result of this dwarfism, and some scientists believe it could actually be a reduced Homo erectus. The majority opinion today is that they were already so small when they reached Flores (such as the  Australopithecus from whom evolved), and modern features are due to convergent evolution with H. sapiens. Unfortunately it has not been able to extract DNA in good condition to put them in the phylogenetic tree for sure.

How did they get to Flores? They had a language, art and cultural expressions? Did they get in contact with our species? They were extinct due to a volcanic eruption? Who made the other ancient tools previous to H. floresiensis? The debate and the unknowns remain open.

REFERENCES

 

You are a bit Neanderthal

Neanderthals are perhaps the better known ancestor for the general public, as like as Australopithecus afarensis (Lucy). The classical view of them, a rough, coarse, crude, unintelligent beings, is still alive in the popular imagination (even “Neanderthal” is used as an insult), but in recent years the research tells us they were not like this. Discover in this article who they were and why you’re a bit Neanderthal.

WHAT WAS A NEANDERTHAL LIKE?

Homo neanderthalensis was the first hominin fossil discovered and currently we have hundreds of fossil specimens of all ages, so is the best known fossil hominin. Got its name from the Neander Valley (Neanderthal in German), a valley near Düsseldorf.

distribución geográfica neandertal
Geographical distribution of the Neanderthals. Image by Ryulong

They lived in Europe (including Siberia) and southwestern Asia,  350,000-28,000 years ago (40,000 according to some sources), an era marked by glacial cycles. They existed in the world longer than us, Homo sapiens.

Neanderthals had various adaptations to cold, as robustness and less height than H. sapiens, and a wide nasal cavity.

Comparación del cráneo de sapiens i neanderthalensis. Cleveland Museum of Natural History. Foto de Matt Celeskey.
Skull comparison between sapiens and neanderthalensis. Cleveland Museum of Natural History. Photo by Matt Celeskey.

Skull highlights its size, with an average capacity of 1,475 cm3, somewhat higher than the modern human skull, and is more elongated backwards (protrusion or occipital bun). Also it is observed easily a powerful supraorbital torus (bone above the eye sockets). Their pelvis was wider than ours and had shorter legs.

homo neanderthalensis, hombre de neandertal, neanderthal, american museum of natural history, amnh
Recreation of a skeleton of H. neanderthalensis. American Museum of Natural History. Photo by Mireia Querol

HOW THEY LIVED?

FEEDING

Neanderthals were skilled and selective hunters, they faced large animals (as witnessed from their injuries, some fatal) and used hunting strategies like the populations of Homo sapiens that arrived in Europe after them. They were seasonal hunters due to seasonal climate (basically reindeer in winter, deer and wild boar in summer). So their diet was based on meat, but near the coast also ate molluscs such as mussels, which were boiled to open them. It is likely that practiced cannibalism. Also captured marine mammals such as monk seals and dolphins stranded, and also ate cooked cereals.

UTENSILS

Homo neanderthalensis had a lithic industry (stone work for constructing tools) called Mousterian, also associated with other species such as H. heidelbergensis and Homo sapiens that requires great skill and planning. In some deposits it has been found composite tools using adhesives.

Neandertal con ornamentación de plumas. Reconstrucción de Fabio Fogliazza.
Neanderthal with feather ornamentation. Reconstruction by Fabio Fogliazza.

There are no remains of clothes, but is likely that they used fur to cover them given the climatic changes that they faced.

In Spanish caves perforated shells were found with traces of pigments, suggesting that they were used as dishes for body painting or dyeing fur. It is suggested that perhaps they were the first to make cave paintings, contrary to the belief that we are the only ones who did it. They also carved bone and used feathers as personal decoration. All this suggests some sort of symbolic thought, associated until recently as an exclusive feature of Homo sapiens.

pintura rupestre, manos, arte rupestre, pinturas rupestres más antiguas, pinturas neandertales
Cave paintings of hands (“Groups of hands”) and red disks in El Castillo cave, Spain. They are the oldest in Europe (41,000 years) and maybe Neanderthals painted it, rather than sapiens as previously thought. Photo: Science.

SOCIETY

Neanderthals are believed that lived in family groups, although recent studies suggest that females would move to other families when they reached adulthood, while adult men remained with the original family.

entierro, neandertal, neanderthal, compasion, autoconciencia
Neanderthal showing compassion to a dead partner. Recreations by Elisabeth Daynès, CosmoCaixa Barcelona. Photo by Mireia Querol

One of the most important features of the Neanderthals is that they were probably the first human ancestors that buried their dead, which shows an awareness of the individual self and their peers, plus some symbolic or abstract thought as mentioned above. This increased the survival of individuals and made stronger social bonds, and also helped other dependent people such as elderly and sick fellows (as the old man from La Chapelle-aux-Saints). Their life expectancy was about 40 years.

DID THE NEANDERTHALS TALK?

Another unanswered question, though are reaching strength opinions of some scientists as Juan Luis Arsuaga, thanks to the remains of the site with more fossils of Homo in the world, La Sima de los Huesos (Burgos). Neandedrthals could have an oral language, against the widespread thinking so far that they had a communication based in grunts. In addition to the anatomical language adaptations, the Neanderthal DNA contains the FoxP2 gene, related to speech in H. sapiens.

neanderthal
Recreation of a Neanderthal camp. Neanderthal Museum in Krapina, Croatia.

NEANDERTHAL EXTINCTION

The extinction of Neanderthals is one of the most controversial debates in paleoanthropology. They disappeared 28,000 years ago, after the arrival of anatomically modern humans in Europe about 60,000 years ago. A time ago extinction was associated with their lower intellectual capacity, but we have seen that did not have to be this way, since they were much like us. Inability to adapt to climate changes? Less reproductive capacity? More infant mortality? Less efficiency for resources or hunting? Direct wars? Imported diseases? Or … maybe sex?

HYBRIDIZATION BETWEEN H. SAPIENS  AND H. NEANDERTHALENSIS

Refused for a long time, we now know that our species reproduced with Neanderthals when they were about to be genetically incompatible (100,000 years ago), because they coexisted between 2,600 and 5,400 years ago and left fertile offspring. So much so, that the Neanderthal genome accumulated by all living human beings is 20%, although the percentage in an individual -without african roots-  is from 1 to 3%.

In June 22nd was published in Nature the discovery of a jaw in Romania of an anatomically modern Homo sapiens (40,000 years old) containing between 6 and 9% of Neanderthal DNA, which implies that their neanderthalensis ancestry was only 4 or 6 generations back in his pedigree.

pelirrojo, redhead, neanderthal
Redhead Homo sapiens and recreation of H. neanderthalensis. Photo by Science

So another possible explanation for their extinction is due to these reproductive crossings. Homo sapiens were more numerous, which could have caused that the Neanderthal genes were diluted” over thousands of generations. This is known as extermination by hybridization.

WHAT IMPLICATIONS WE HAVE BEING A BIT NEANDERTHALS?

It is believed that Neanderthals genes brought us some advantages, as some characteristics of the skin and hair, such as color and thickness, which could help our species to colonize cooler areas. In fact some Neanderthals could be light-skinned and redheads.

But some diseases can be associated to that heritage: increased risk of biliary cirrhosis, lupus, diabetes, Crohn’s disease and even difficulty in quitting smoking (smokers: not worth using it as an excuse).

In short, it is exciting to think that we lived and even mated with a species so similar to ours and that somehow, still exist in each of us. We may not be as special as we thought.

Currently we are the only representatives of the genus Homo, but in ancient times it was not. Can you imagine a world where you would meet  a Neanderthal  in the street and tell them good morning”?

 

neanderthal, suit, traje, camisa, nenadertal
Neanderthal in suit. Photo: Neanderthal Museum/H. Neumann

REFERENCES

MIREIA QUEROL ALL YOU NEED IS BIOLOGY

Flowers wearing turban, the Tulip fever

The spring beginning has allowed some of you to enjoy the beautiful colours of those flowers that have already bloomed. This time I’m going to talk about one of the most colourful, simple, but wonderful flowers you probably already will have had the opportunity to observe in many gardens or in nature. It is the tulip. Besides introduce you this plant, in this article I will make a more detailed description of its morphological parts. I think it’s a good example to start learning vocabulary, because its structure is quite clear and simple. Therefore, if you are interested in learning some technical vocabulary, now it’s a perfect chance. But, do not think I’m just going to talk about the technical aspects, because reading this article you will also be able to learn the history behind the tulips. And as you will see, these flowers caused a good fever!

tulipes
Artistic image of several tulips (Photo taken by Adriel Acosta).

 INTRODUCTION

The tulips (Tulipa sp.) are flowers that when are closed seem a turban. This plants have been very popular and well-known for very long time, because of its high ornamental interest.

Its genus is distributed in the central and western Asia, in the Mediterranean and in Europe. It is known that its origin belongs to the centre of Asia and, from there, their distribution has been expanded naturally and by human actions. And, although about 150 species are known in the nature, human intervention has greatly increased the species list. Caused both by hybridization (forcing the offspring of two interesting species) and by selective breeding (choosing the offspring which has more value).

Tulipa_cultivars_Amsterdam
Tulip crop in Amsterdam (Photo taken by Rob Young). 

 THE TULIP FEVER

As already mentioned above, tulips are one of the most ornamental plants used, both in decoration as in landscaping. And while the tulip crop is rather old, the boom occurred in Europe during the seventeenth century. Giving rise to what is known as Tulip mania or the Tulip fever. In those moments, especially in Netherlands and France, a high interest in the cultivation of these plants awoke. The fever was so great that people were selling goods of all kinds to buy tulip bulbs, even reaching up to sell the most valued as the house or farm animals.

The cause of this was originated in the Netherlands, where the single-coloured tulip bulbs were being sold at that time. But afterwards, the Eastern bulbs that give rise to flowers with variegated colours appeared. And they were very attractive. Although the cause was uncertain in that moment, it was known that if a single-coloured bulb touched other marbled-coloured bulb, the first one would turned into a marbled-coloured bulb. This caused the tulip’s price began to increase and soon after occurred the first speculative bubble in history.

Nowadays, we know that the cause is due to a virus which is transmitted from some bulbs to others; this virus is known as Tulip breaking virus.

Semper Augustus Tulip 17th century
Anonymous gouache on paper drawing, 17th century, of the “Semper Augustus”. A representation of one of the most popular tulips which was sold at record price in Netherlands (Public Domain).

MORPHOLOGICAL CHARACTERS

 The plant

 Tulips are geophytes, that is, they have resistance bodies underground to survive during unfavourable seasons, the winter. These organs are bulbs, which have been used on crops to preserve these plants.

Its leaves are linear or linear-lanceolate, i.e., they are long, narrow and acute. Parallel venation can be observed on its leaves, so a nerve is by side other and with the same direction. Their arrangement is usually in rosette: this means that the leaves are born agglomerated in the bottom of the plant above the bulb, and at the same level. Even so, you can sometimes see some leaves along the stem, cauline ones. These are sessile, without petiole, and wrap a little the stem.

To cultivate tulips, we can use their bulbs or fruits. These seconds are capsules, a dried fruits, opened due the action of some valves. At first, the seeds are hooked inside these capsules and then are released and distributed on the environment.

20150329_165102[1]
Tulip (Photo taken by Adriel Acosta).

The flowers

Tulips appear in early spring, due they are plants adapted to very dry Mediterranean climate or cold areas.

As you have seen, the flowers are solitary or appear to 3 gathered in one stem. They are usually large and showy, hermaphrodite, therefore, they have both male and female reproductive organs, and are actinomorphous, that is, they can be divided symmetrically for more than two planes of symmetry.

These flowers have 3 inner tepals and 3 external that are free among them, without being bound or fused. We talk about tepals when the sepals (calyx pieces) and petals (corolla parts) are similar between them. In this case, the tepals are petaloid, because they adopt typical colours and shapes of the petals.

In the inner part of the flower, we can see 6 stamens divided equally into 2 whorls; being these two closely spaced between them, so they seem to arise from the same point. And right in the centre, surrounded by these stamens, there is the gynoecium, female part of the flower. This gynoecium consists of the ovary and 3 stigmas attached to this directly. The stigmas are this part of female reproductive organs where it should arrive pollen to fertilize the ovaries.

part tulipa
Parts of tulip flower: 1. Sepal, 2. Petal, 3. Stamen, 4. Female reproductive organ (ovary and 3 stigmas) (Photo taken by Adriel Acosta).

 As you have seen in this article, some flowers have caused curious stories and a great impact on our society. Also, you have had the opportunity to observe in detail the tulip’s structure. One more time, I wish you liked it.

Difusió-anglès

REFERENCES

  • A. Aguilella & F. Puche. 2004. Diccionari de botànica. Colleció Educació. Material. Universitat de València: pp. 500.
  • Bolòs, J. Vigo, R. M. Masalles & J. M. Ninot. 2005. Flora manual dels Països catalans. 3ed. Pòrtic Natura, Barcelona: pp. 1310.
  • Notes of Phanerogamae and Applied Plant Physiology, Degree of Environmental Biology, Ambiental, UAB
  • F. Schiappacasse. Cultivo del tulipan. http://www2.inia.cl/medios/biblioteca/seriesinia/NR21768.pdf
  • Fundación para la Innovación Agraria; Ministerio de Agricultura. 2008. Resultados y Lecciones en Tulipán. Proyecto de Innovación en XII Región de Magallanes. Flores y FOllajes/ Flores de corte (11).