ES2220157B1 - GROUP OF PLANTS, METHOD FOR PLANTING TREES AND METHOD FOR CULTIVATING PLANTS. - Google Patents

Abstract

Group of plants, method for planting trees and method for growing plants. Inhibition of damage to forests caused by ultraviolet rays. This is done by the selection by triploid or tetraploid cochromine type of chromosomes and by repopulation to form a group of plants resistant to ultraviolet rays creating an artificial forest.

Description

Forest planted by man, method for plant trees and method of growing plants.

Background of the invention

The present invention relates to a technique of  plant cultivation, such as tree cultivation, and more specifically to a selective tree cultivation technique young or similar that is resistant to ultraviolet rays with object of obtaining vegetation, such as that of a forest, reliably resistant to ultraviolet rays.

Lately, the problem of lightning Ultraviolet on earth is the subject of debate. Must be especially to the fact that in recent times it is destroying the ozone of the stratosphere surrounding the earth, with which ultraviolet rays reach the earth from the outside and exert their bad influence on terrestrial biology.

For example, in relation to the human body, it  reports on the increase in conditions associated with lightning ultraviolet, and it is confirmed that the increase in cancer cases of the skin is due to an increase in radiation from the ultraviolet rays. Therefore, it is estimated that the damage derived from Ultraviolet rays results in the presence of active oxygen. Niwa, one of the present inventors, has studied its properties Pharmacological and biochemical conditions of various conditions caused by active oxygen over several years.

Summary of the Invention

Actual damage from lightning Conventional ultraviolet have been studied primarily, and as a primary topic, in relation to its effects on the body human. Moreover, the present inventors have reached the conclusion that ultraviolet rays have also caused a considerable damage to forests.

It has been determined that the greatest damage to forests  It comes from acid rain. For example, in the district of Schwarzwald, Germany, a report on the considerable damage caused in an extensive area where large number of trees perished due to acid rain. Oxides  acidic, such as sulfur-containing oxides derived of industrial combustion, they dissolve in raindrops to become acid rain.

Research on these effects was repeated.  on several occasions by the present inventors, who visited the Damaged forest and obtained extensive results.

The present inventors observed the damages of the forest in the course of several visits made to various places of the same, in different districts. After a detailed investigation of the conditions observed, the present researchers determined that there are possibly others factors, in addition to acid rain, strongly related to abnormal phenomena of withered trees found in the forest. Such damage could be due to ultraviolet rays, identified as the main factor related to damage to forest.

There are not many previous studies on harmful effects of ultraviolet rays on forests. How result of the effects observed by the present inventors in In relation to ultraviolet rays, it is necessary to take measures urgent practices aimed at inhibiting such damage to forests. Due to the loss of ozone from the atmosphere and the increase in ultraviolet radiation worldwide, measures must be taken urgent to inhibit the damage caused by ultraviolet rays,  focusing on the various factors that cause such damage. Yes no measures are applied in this regard, forests around the world are they will wilt due to ultraviolet rays, and their global disappearance

The object of the present invention is to prevent  damage to forests caused by ultraviolet rays.

Man-planted forests according to the present invention are characterized by the cultivation of seed plants endowed with polyploids ( tasusei , in Japanese). The chromosome number of seed plants is triploid or tetraploid. The plants are coniferous. Such coniferous plants are selected from the group that includes Japanese cedar and cypress. The largest number of seed chromosomes is obtained through reproduction and mutation.

The tree cultivation method of the present invention is characterized in that it comprises the steps of selecting seed plants with polyploid chromosomes and then planting them. The number of chromosomes in plants must be triploid or tetraploid. The plants are coniferous. Such coniferous plants are from Cryptomeria japonica D. Don and Chamaecyparis obtusa Endl . The largest number of seed chromosomes is obtained through reproduction and mutation. The plant cultivation method of the present invention is characterized in that it comprises the steps of forming an artificial vegetation considerably resistant to ultraviolet rays compared to the vegetation constituted by triploid plants - by selective cultivation of plants whose chromosome number exceeds the number basic and intrinsic of the species.

In this specification, "polyploid" ( tasusei , in Japanese) defines those species with three times more chromosome sets - that is, the number of fundamental chromosomes multiplied by three. In addition, it should be borne in mind that the haploid or diploid normally observed in plants is not included in the scope defined by said polyploid.

Description of the preferred embodiments

Several examples are described below and details of the realizations.

Seed plants used in forests man-planted, the method of planting trees and the method for growing plants of the present invention include aspects of resistance to ultraviolet rays (hereinafter, "UV resistance". Afforestation, in the case of forests planted by man, it is done using seed plants with this property of resistance to ultraviolet rays.

Once the seed plants that have such UV resistance property are selected, a standard for the selection of polyploid chromosomes ( tasusei ) acquired by said plants is adopted. The term polyploid used herein defines seed plants with chromosome numbers greater than diploid.

This study has been proposed, for the first time, by the present inventors. The trees with chromosomes polyploids possess a resistance against ultraviolet rays higher than normally observed in triploid trees. In the man-planted forests formed by the aforementioned plants UV resistant seed, an obvious inhibition of damage caused by ultraviolet rays on trees with triploid chromosomes. Abnormal phenomena refer to effects such as standing withering trees, perennial leaves They dry out and deciduous leaves fall out of time. It is estimated that these abnormal phenomena, produced by ultraviolet rays, They can be inhibited.

Several are described in detail below.  types of trees and several examples of the effectiveness of the invention.

Example

In the example, leaves were collected from plants  described in sections (1) to (3) below include, subjecting them to lightning resistance experiments ultraviolet (resistance W).

(1) Rice ( Oryza sativa )

Oryza sativa leaves were used for comparative purposes of the samples. Oryza sativa is a monocot plant well known for its resistance to ultraviolet rays. Samples of Oryza Sativa leaves used in the experiment were obtained in the city of Tosashimizu, Kochi Prefecture, in Shokoku, Japan. The leaves of Oryza Sativa , from five different areas of the city of Tosashimizu, were subjected to measurement of enzyme activity by the procedure described below.

(2) Osmunda ( Osmunda japonica Thunb )

For the experiment, Osmunda japonica Thunb leaves were collected that grow spontaneously in Tosashimizu, Japan. It is known that the Osmunda japonica Thunb , a fern, is a silver whose evolution is earlier than that of other plants. Many reports indicate that the Osmunda japonica Thunb adapts to environmental conditions thanks to the fact that they have acquired the polyploidy multiplier of chromosomes of the species.

(3) Gymnosperm, such as black pine, etc.

\hbox{a 4.}

Being a plant sensitive to ultraviolet rays, gymnosperm leaves were included in the experiment. Tables 1 to 4 of this example include the Kuromatsu tree (whose Latin name is Pinus Thunbergii Parl ), the sugi tree (whose Latin name is Cryptomeria japonica D. Don ) and the Hinoki tree (whose Latin name Chamaecyparis obtusa Endl .), which grow spontaneously in the mountains of Japan and whose leaves were used for the experiment. Said samples of black pine, cedar and cypress leaves were obtained from the sites reflected in tables 1

 \ hbox {a 4.} 

The black pine (hereinafter referred to as "pine control black ") sensitive to common UV rays and pine UV resistant black (hereinafter referred to as "pine UV resistant black "), described separately in Table 1, underwent tests of SOD activity and activity GSH-Px, and to tests to determine its level of lipid peroxide.

In addition, and as reflected in Tables 5 and 6, triploid species and species measurements were made tetraploids derived from crosses and mutations.

TABLE 1 Comparison of levels of lipid peroxide and SOD and glutathione peroxidase (GSH-Px) activities between Japanese black pines ( Pinus Thunbergii Parl .) UV resistant and Japanese black pines ( Pinus Thunbergii Parl .) UV sensitive

one

\hskip1cm

* P < 0,01 entre pinos negros resistentes y sensibles a UV

 \ hskip1cm 

* P <0.01 among resistant and UV-sensitive black pines

TABLE 2 Comparison of lipid peroxide levels and SOD and GSH-Px activities in Japanese cypress ( Chamaecyparis obtusa Endl .) Triploids

2

TABLE 3 Comparison of lipid peroxide levels and SOD and GSH-Px activities in additional units of Japanese cypress ( Chamaecyparis obtusa Endl .)

3

       \ dotable {\ tabskip \ tabcolsep # \ hfil \ tabskip0ptplus1fil \ dddarstrut \ cr} {
 \ begin {minipage} {130mm} + P <0.001 between tetraploids of
Japanese cypress control units (Chamaecyparis obtusa
Endl.) And diploids or triploids of Japanese cypress
(Chamaecyparis obtusa Endl.), And between tetraploids of
Japanese cypress control units (Chamaecyparis obtusa
Endl.) And diploids or triploids of cypress control units
Japanese (Chamaecyparis obtusa
Endl.). \ End {minipage} \ cr}
    

TABLE 4 Comparison of lipid peroxide levels and SOD and GSH-Px activities in control units of Japanese cedar ( Cryptomería japan D. Don ), rice leaves ( Oryza Sativa ) and osmunda ( Osmunda japan Thunb )

4

       \ dotable {\ tabskip \ tabcolsep # \ hfil \ tabskip0ptplus1fil \ dddarstrut \ cr} {
 \ begin {minipage} {130mm} \ pm <0.0001 between tetraploids and
Japanese cedar control unit triploids (Cryptomería
Japan D. Don.) \ end {minipage} \ cr
 \ begin {minipage} {130mm} \ NAK P <0.00001 between
Tetraploids and triploids of Japanese cedar control units
(Japan Cryptomería D.
Don.). \ End {minipage} \ cr}
    

As indicated in Table 1, the Pinus Thunbergii Parl . UV resistant (W resistant) were compared with the Pinus Thunbergii Parl . sensitive to ultraviolet rays (control units) in relation to SOD activity, glutathione peroxidase activity (GSH-Px) and lipid peroxide levels.

In Table 2, the triploids of Chamaecyparis obtusa Endl . they were compared with the triploids of Chamaecyparis obtusa Endl . in relation to lipid peroxide levels, SOD activity and GSH-Px activity.

In Table 3, the diploids, triploids and tetraploids of control units of Chamaecyparis obtusa Endl . they were compared in relation to individual SOD activity, GSH-Px activity and lipid peroxide levels.

Table 4 compared the SOD activity, the GSH-Px activity and the lipid peroxide levels of the diploids, triploids and tetraploids of control units of Cryptomería japan D. Don , leaves of Oryza Sativa and Osmunda Japan Thunb . The comparison between table 4 and tables 5 and 6 is described below.

Extracts of diploid, triploid and tetraploid leaves from Cryptomería japan D. Don and Chamaecyparis obtusa Endl . they are compared in tables 5 and 6, as described below. In table 5, column of control samples, the brand used to describe the sample size of 0.6 gr / ml is called "x1", that of 0.3 gr / ml is called "x2", that of 6.0 gr / ml is called "x10". Table 6 includes information on Oryza Sativa sheets, for comparative purposes.

The individual enzymatic activity and the level of lipid peroxide of the plants described in sections (1) to (3) were tested. The test elements used in the measurement of enzymatic activity consisted of SOD activity, glutathione peroxidase activity (GSH-Px) and catalase activity. The results obtained are included in tables 1 to 4. The amounts appear in the form of ± SD. On the contrary, it was totally impossible to detect the catalase activity of plants other than Oryza Sativa , so their results are not listed in Tables 1 to 4.

In relation to the quantitative analysis of lipid peroxide level, the level in question was measured using the TBA reaction method, which consists in converting the docohexanoic acid in a substance reactive to TBA.

Homogenates of plant leaves Individuals described above, used as samples, are They prepared as follows. The collected leaves are they pulverized in a mill. The homogenates of the leaves of the individual plants described above were used as samples. The procedure for preparing said homogenates of sheet is as follows. The powder is suspended in a solution of 95% ethanol for the lipid peroxide formation test or in a physiological saline solution for any other test that does not be of lipid peroxide formation. In both cases, the concentration is 60 mg / ml. The suspension solution is subjected to sonication (15 W, for 15 seconds).

After sonication, the solution is centrifuge at 1,300 g, as shown in figure 1. Once performed the centrifugation, the supernatants are divided into two parts A part is used in the preparation of samples for Determine the level of lipid peroxide. The other is passed through of a 45 µ filter. A part of this filtering is used as sample for the SOD activity test, and the rest as samples for catalase activity tests and GSH-Px.

Quantitative SOD Analysis

The mixture indicated above is added to 0.2 ml of the test mixture containing xanthine-xanthine oxidase to produce O2 -. The resulting amount of O2 - is determined by the amount of reduction of ferricitochrome c (type III) obtained by absorbance 550 nm optics, measured with a spectrophotometer (Beckman).

The amount of SOD contained in the sample is defined as the amount of reduction obtained by reducing cytochrome 50% c One unit of SOD is expressed in units / mg of protein. In this analysis, the sample itself not only inhibits the reaction on cytochrome c without the mediation of O2 - but reduces directly a small amount of cytochrome c.

Using these factors, the amount was calculated Actual COD by means of the following formula:

unit = {a- (b-c)} / (a / 2)

in where:

a = absorbance obtained by the addition of only Xanthine oxidase.

b = absorbance obtained by homogenates of the leaves in the presence of xanthine oxidase.

c = absorbance obtained by the addition of only leaf homogenates before the addition of xanthine oxidase

Catalan activity test

The catalase activity was determined on the basis of the rate of reduction of hydrogen peroxide (H 2 O 2). The rate of reduction was measured for 12 to 30 seconds, in the presence of samples of leaf homogenates, using a spectrophotometer at 240 nm.

The activity is expressed through the following  formula:

K = (2.3 / 18) X log (A1 / A2)

A1 = Absorbance measured for 12 seconds at 240 nm in test mixture with a total volume of 3.1 ml, consistent in 10 mmol / L of H2O2 in a 3 ml solution of 50 mmol / L of potassium phosphate buffer solution and 0.1 ml of homogenate leaves.

A2 = Absorbance measured for 30 seconds at 240 nm in said test mixture.

In this test, the peroxide substrate of hydrogen was replaced by sodium perborate due to the possibility of hydrogen peroxide causing instability. In this case, a solution of 0.002 to 0.05 ml of Leaf sample was suspended in 2.8 ml of buffer solution (pH 7.4) 0.5 mol / L potassium phosphate, keeping the mixture in preincubation at 30 ° C for 5 minutes. Then it was added a 0.2 N solution of sodium perborate to the pre-incubated mixture, starting the reaction. Information was recorded during 2-3 minutes, at 220 nm.

GSH-Px activity test

For the measurement of the GSH-Px, he used the method of Lawrence and Burk, of connection with oxidation of NADPH from gutathione reductase. The oxidation power of NADPH it was measured at 37 ° C, using a spectrophotometer with a length 340 nm wave

The test mixture consisted of 50 mmmol / L of potassium phosphate buffer solution (pH 7.0), 1 mmol / L acid ethylenediaminetetraacetic acid, 1 mmol / L NaN3, 0.2 mmol / L glutathione, 2 glutathione reductase units, and 1.5 mmol / L of 10 mmol / L cumene hydroperoxide solution tert-butylperoxide.

To this mixture was added homogenate of the leaves, obtaining a solution volume of 1.0 ml. Activity Enzymatic measured by this method equals the amount of oxidized NADPH per minute. The activity is represented in units / mg of protein, according to Lowry's method.

Lipid Peroxide Assay

Polyunsaturated fatty acid 4,7,10,13,16, 19-docosahexanoic was diluted 200 times in ethanol 94% To the test tube containing this substrate were added various concentrations of homogenates before subjecting it to ultraviolet radiation In the experiment homogenates were tested with concentrations of 0.6, 3.0 and 6.0 mg / ml. After the radiation, the solution was analyzed by means of fluorospectrometry to measure the existing amount of TBA (acid thiobarbituric). Wavelengths were set at 515 nm for excitation and 553 nm for emission in a fluorospectrophotometer Hitachi F-2000.

The results appear with the average indication S.E. The amount of TBA substance was adopted as a measure of lipid peroxide. In addition, a difference analysis was performed significant through a Student t test. The t test result It demonstrates the significant difference between the two groups.

Identification of the basic numbers of chromosomes

The ploids of the aforementioned trees were determined through cytological analysis of cells from root tips or cell flow cytometry analysis of leaves.

The debate on tables 1 to 6 was carried out as follows. As shown in Tables 1 to 4, the leaves of Oryza Sativa and Osmunda Japan Thunb (ref. Table 4) reflected a SOD activity apparently higher than those of gymnosperm plants, such as the Pinus Thunbergii Parl ., The Japanese Cryptomería D. Don and the Chamaecyparis obtusa Endl . The GHS-Px of the leaves of Oryza Sativa stands out , which reflected a high activity of 1384 units / g, and the SOD, whose great activity reached 2941 units / g.

As can be seen in Table 4, the SOD activity of the Oryza Sativa leaves is superior to that of the individual trees under test (except for the Cryptomería japan D. Don , which is a tetraploid plant). In some cases, the greater SOD activity is equivalent to 100 times the activity of the common trees under test. In Table 4, the leaves of Osmunda Japan Thunb reflect a high activity, reaching 1394 units / g.

As for the gymnosperm plants tested, and as shown in Table 1 (P <0.01), it is seen that the Pinus Thunbergii Parl . they have ultraviolet resistance properties, their SOD activity apparently being higher than that of the Pinus Thunbergii Parl . common. For example, the SOD activity of the Pinus Thunbergii Parl . that possess properties of resistance to ultraviolet rays is 413 ± 56 units / g on average, a much higher value than that of the Pinus Thunbergii Parl . common (240 ± 37 units / g on average).

From the previous results it follows that  trees that have lightning resistance properties Ultraviolet have a high SOD activity. This would indicate that the SOD activity is involved in lightning protection ultraviolet.

When comparing the SOD activity of the triploid tree species and the tetraploid tree species, as shown in Tables 2 to 3 and especially with reference to the individual trees, the tratraploid tree species exhibited a greater SOD activity than the species of triploid trees. The control tree of Japan's Cryptomería D. Don (sample of Kamikawa cedar, Cr-38c) that appears in table 4, revealed a SOD activity much greater than that of common trees. In said Cryptomería japan D. Don , the apparent increase in SOD activity was detected in trees that had the tetraploid chromosome. In contrast to this, neither diploid trees nor triploids reflected an apparent increase in SOD activity.

In addition, regarding the peroxide level lipid, it has been shown that cedars with small amounts of resin have a capacity of formation of lipid peroxide considerably smaller than that of other trees.

No difference in GSH-Px activity was found between the Pinus Thunbergii Parl . resistant to ultraviolet rays and the Pinus Thunbergii Parl . ultraviolet-sensitive commons included as control units in the table (ref. table 1, P> 0.05). This could indicate that there is no relationship with GSH-Px enzymes resistant to ultraviolet rays.

TABLE 5 Comparison of SOD activity and the effect of lipid peroxidation of leaf extract among Japanese trees 2X, 3X and 4X

5

       \ dotable {\ tabskip \ tabcolsep # \ hfil \ tabskip0ptplus1fil \ dddarstrut \ cr} {
 \ begin {minipage} {130mm} * x1 means 0.6 gr / ml as a unit, and
the sample column added indicates the multiple of the
drive \ end {minipage} \ cr}
    

TABLE 6 Comparison of SOD activity and the effect of lipid peroxidation of leaf extract among Japanese trees 2X, 3X and 4X

6

       \ dotable {\ tabskip \ tabcolsep # \ hfil \ tabskip0ptplus1fil \ dddarstrut \ cr} {
 \ begin {minipage} {130mm} * x1 means 0.6 gr / ml as a unit, and
the sample column added indicates the multiple of the
drive \ end {minipage} \ cr}
    

The formation of lipid peroxide increased with a  low concentration of homogenate (for example, 0.6 mg / mi). A high concentration of homogenate inhibits peroxide formation lipid This is clearly shown in Tables 5 and 6.

As can be seen in Tables 5 and 6, the comparison between samples of tetraploid trees indicates clearly that samples with high SOD activity have a strong inhibition effect in the presence of low samples concentration. This might suggest that the SOD activity plays a role in inhibiting the formation of lipid peroxide as cytotoxic element. This means that, the more SOD activity, increased inhibition of lipid peroxide formation.

For example, the samples of Cryptomería japan D. Don Nº S-26, -30, -32, -34, -36 and -38 of table 5 and the samples of Chamaecyparis obtusa Endl . No. H-6, -9, -10 and -11 of Table 6 have an SOD activity that exceeds 1000 units / g. Among these samples, nine of them, Nos. S-26, -30, -32, -34 and H-6, -9, -10 reflected a lower formation of lipid peroxide (as indicated in column x1 of the table 5) by adding samples with a lower concentration than the controls.

In the case of the Oryza Sativa leaf sample, the formation of lipid peroxide was inhibited irrespective of the low concentration (see column x1 of table 5) or the high concentration (see column x10 of the table) of homogenates Therefore, the SOD activity was as high as the GHS-Px activity and the catalase activity. Among the leaf samples tested, the Oryza Sativa leaf reflected the highest level of lipid peroxide formation. In other samples, other than those of Oryza Sativa leaves, no catalase activity was detected, so these data have not been collected in the tables.

On the contrary, the greater the activity  SOD, the stronger the effects of antioxidation. SOD It plays an important role in the removal of active oxygen.

When considering factors related to formation of active oxygen, the presence of rays is not ruled out ultraviolet. In recent years, the destruction of the ozone layer has generated an increase in ultraviolet radiation that reaches the land. Based on this evidence, a apparent increase in oxygen radicals. Like the humans, trees suffer the effects of exposure to ultraviolet rays and high concentrations of radicals oxygen.

However, the experimental results of the  present inventors point out that the SOD activity is high in trees that possess ultraviolet resistance, so it is estimated that this aspect constitutes a function destined to its protection against environmental damage with the use of antioxidant agents such as SOD that found in its leaves. Until now there was no report concerning the self-protection of trees, through their own antioxidant agents, against the damage caused by the environment. This is the discovery of those present inventors

The antioxidant capacity of pine trees resistant to ultraviolet rays (hereinafter referred to as "UV resistant") was compared to that of common pine trees. Reports on the induction of GHS-Px activity in response to oxygen radicals were obtained. However, the experiments carried out by the present inventors did not provide any results on apparent changes in GHS-Px activity in parallel to the activity.
SOD

Regarding catalase, there is a report related to the production of catalase induced by tree oxygen radicals. However, this experiment detected catalase only in the leaves of Oryza Sativa . Terpene and other elements of homogenates are thought to inhibit activation.

It is a widely known fact that, like that enzymes (such as superoxidase), substances from low molecular weight (such as vitamin C in its function of oxidizing agent) play an important role in the lives of trees. In the experiment of this invention the antioxidant function of low molecular weight compounds, although the SOD activity was studied. The conclusion was that the SOD, a strongly antioxidant enzyme, is produced in the leaves of trees to protect them against radical excesses of oxygen.

As a result of the previous analysis, and as they have  discovered the present inventors, when considering the relationship between SOD activity and the chromosomes of plants, trees of the triploid and tetraploid type, and particularly of the type tetraploid, reflect a high SOD activity. It has not been analyzed the relationship between SOD activity and tree based ploids in the studies of other specialists. In accordance with the above previously, the results obtained by the present Inventors have not been evaluated at all.

When considering the relationship of the SOD activity with  the polyploid of the chromosomes and the increase of the locus of the SOD chromosome coding, there is an increase in the existing amount of SOD. This consideration can be affirmative. in relation, for example, to human beings. It has been determined that Down syndrome has three chromosomes (trisomic) containing Cu, Zn and SOD locus, which produces a 50% increase in the amount of SOD.

Despite the increase in SOD activity individually detected in triploid and tetraploid trees at through the experiments carried out by those present inventors, no potentiation of the level of SOD This means that an elucidation of another type. However, the conclusion is that a increased positions of SOD chromosomes in samples Individuals tested.

It is estimated that the leaves of Oryza Sativa are endowed with resistance to ultraviolet rays, that their SOD activity is greater than that of the Japan Don Cryptomería , the black pine and the control units of Chamaecyparis obtusa Endl ., And that, in addition , have a great activity GHS-Px, superior to that of the mentioned trees. In addition, catalase, almost undetected in these trees, did reflect a detectable level in the leaves of Oryza Sativa .

This means that, through the capacity that provides the strong enzymatic activity that trees possess in response to oxygen radicals and ultraviolet rays from the environment, monocot plants can acquire high adaptability to defend against ultraviolet rays  enviroment. For example, ferns are more plants primitives than gymnosperms and angiosperms. It is a fact well known that ferns can achieve great adaptability by multiplying its number of chromosomes to convert it into polyploid Recently, there has also been a greater Adaptability of bacteria against disturbances environmental through the acquisition of functions polyploids

In addition, the experiments described above demonstrate that the Japanese Don Cryptomería D. and the Chamaecyparis obtusa Endl ., Which have ultraviolet resistance properties, have triploid and tetraploid chromosomes.

Even gymnosperm plants sensitive to ultraviolet rays can increase your SOD activity by multiplication of your chromosome set in order to protect yourself from themselves against the longest times of exposure to lightning ultraviolet and oxygen radicals. Apparently, this means that species have a considerable capacity to adaptation.

On the other hand, it is possible that the black pine resistant to ultraviolet rays increases its SOD in order to protect itself without this being related to polyploid chromosomes, unlike the common black pine. It is also estimated that the black pine resistant to ultraviolet rays does not adopt an attitude of multiplication of its chromosomes to become a polyploid. As a general rule, it is known that the highest UV-B radiation (where B is the region in which ultraviolet rays are distributed from 280 nm to 320 nm) is not related to oxidation damage of bodies such as those of plants. Therefore, the case of black pine confirms this
hypothesis.

There are two types of polyploid acquisition of chromosomes by plants: one is by natural crossing or artificial and the other is by natural mutation, such as by change of branch or mutation by treatment with colchicine. In this case specifically, the mutation is produced by the procedure of colchicines and by using the mutation of the yolks.

(a) Colchicine procedure

Colchicines are a type of alkaloid. The mutation is induced by the introduction of the seeds of conifer in an aqueous solution of colchicines (0.1% to 0.5%) from 1 to 3 days.

The mutation can also be induced by adding the growing end of the apical bud of the plant to  an aqueous solution of colchicines, so that it can be obtained the tetraploid variant. For example, to get the variant of tetraploid for coniferous seedlings it is preferable to add a aqueous solution of colchicines from 0.2% to 0.4%, 4 or 5 times, one day yes and another no.

(b) By branch change mutation (mutation of the yolks)

The plant that has more chromosomes than fundamental presents a mutation called "change mutation of branch "(mutation of the yolks). This mutation takes place in the nature, although it is unlikely to occur. Thus, This mutation is used as follows: the chromosome tetraploid is obtained through research in a forest that it has resistance to ultraviolet rays without wilting, and selecting seedlings that have tetraploid chromosomes.

This procedure is used because the main feature, which is caused by the mutation, in the fields of agriculture and gardening, as in the case of green tea, apple, banana, tulip orchid, etc.

The Japanese Osmunda Thunb , a fern similar to gymnosperm and angiosperm plants, has already acquired the polyploid condition, as have many types of ferns. It is known that monocoltiledóneas plants, such as the Oryza Sativa , have the same resistance against ultraviolet rays as the Pinus Thumbergii Parl ., The Japanese Don Cryptomería D. and the Chamaecyparis obtusa Endl . The experiments carried out by the present inventors indicate that the leaves of both Osmunda Japan Thunb and Oryza Sativa reflect a stronger SOD activity than that of other trees. Gymnosperm plants, such as black pine, Cryptomería japan D. Don and Chamaecyparis obtusa Endl ., Demonstrate their response to the real environment through the multiplication of their chromosomes with SOD genes, as the present ones have been able to verify inventors

Based on references in the field of agriculture and gardening, it is observed that in many cases there are improvements in various types of vegetation - such as tea plants, apple trees, bananas, tulips and orchids - through the use of polyploids. However, in the case of the Japanese Don Cryptomería D. and the Chamaecyparis obtusa Endl ., It is clear that polyploid species do not develop as well as common species. This value of polyploid species has gone completely unnoticed by the forestry industry sector.
such.

In the control units of Cryptomería japan D. Don and Chamaecyparis obtusa Endl ., The present inventors have found natural triploid species among sterile species through biological cell research. The reports and results obtained by the present inventors reveal that only 41 clones of Japanese cedars and 2 clones of control units of Chamaecyparis obtusa Endl . correspond to triploid species. It is worth mentioning that the "control units" show a good growth rate.

The present inventors have endeavored to cultivate the triploid species using artificial crossing technology. This was done through crosses between triploid and tetraploid species of Cryptomería japan D. Don and Chamaecyparis obtusa Endl ., Respectively. The present inventors have observed that artificial triploid species obtained through artificial crossing developed better than diploid species.

That is, the fact that better growth of triploid artificial crossing species than of common triploid species was confirmed was confirmed. It follows that it is possible and beneficial to use polyploid species to cultivate Cryptomería japan D. Don and Chamaecyparis obtusa Endl .

The lipid peroxide test reflects that the leaf homogenate of tetraploid species with high SOD activity apparently inhibits peroxide formation lipid This evidence could suggest that trees are protected themselves against the damage caused by oxygen radicals introduced by ultraviolet rays through the multiplication of its number of chromosomes.

Japanese cedars - with the exception of tetraploid cedar - that contain a comparatively smaller amount of resin than other trees, reflected a low level of lipid peroxide and a low SOD activity. These results are included in Tables 2 to 4. Because the Japan Don Cryptomería contains a small amount of resin, it is clear that the amount of lipid peroxide that is formed will be low. Therefore, a low SOD activity suffices to treat low lipid oxide formation.

That is to say, it is estimated that Japan's Don Cryptomería does not require a large SOD activity for the treatment of a low level of lipid peroxide, since this cedar does not produce a large amount of said lipid peroxide.
co.

The investigation of the present inventors confirms that trees demonstrate excellent adaptability against the increase in the harmful level of oxygen radicals resulting from the recent acceleration of damage to the ozonosphere. The ferns, which are plants of a more primitive evolution than gymnosperm and angiosperm plants acquired the function polyploid in order to adapt to the disadvantages of environmental alterations through the multiplication of their number of chromosomes

In addition, in needle-shaped leafy plants, such as the Cryptomería japan D. Don and the Chamaecyparis obtusa Endl ., It is observed that their seeds acquired the property of resistance to ultraviolet rays by multiplying their number of chromosomes. This can be seen in the comparison between resistant and non-UV resistant species. It was discovered that especially triploid and tetraploid trees presented this tendency.

Although some natural cross species they showed inadequate growth or became sterile, they confirms that they could achieve good characteristics of growth through artificial crossing.

It was discovered that the means of acquiring the property of adaptability of the Japanese Don Cryptomería D. and of the Chamaecyparis obtusa Endl . It consists of multiplying the chromosonic game to turn it into a polyploid. It can be said that this production of polyploid offspring through artificial crossing constitutes an effective method of protecting trees against the growing tendency of oxygen radicals induced by ultraviolet rays.

This invention is not necessarily limited to previous examples, and other embodiments may be included within the scope of the same as long as they do not depart from their object.

For example, in addition to the aforementioned needle-leaf trees, such as the Cryptomería japan D. Don and the Chamaecyparis obtusa Endl ., The adaptation of broadleaf trees, such as evergreen and deciduous, can be considered. On the other hand, although the trees have acquired their resistance to ultraviolet rays through the multiplication of their chromosome numbers and although the results of this experiment were not obtained from Pinus Thumbergii Parl ., Obviously there could be certain species of Pinus Thumbergii Parl . capable of acquiring resistance to ultraviolet rays by obtaining polyploid media, since there are many types of pine trees.

It is estimated that it is possible to select strong species of plants resistant to ultraviolet rays from of the chromosome games. Selected plants must stand in sunny areas to keep their fort ecological resistance against ultraviolet rays.

Although the above description focuses on the triploid and tetraploid varieties, polyploid trees, such as those with pentaploid and hexaploid chromosomes, They can produce the same results.

In forests planted by man according to In the present invention, such damages are considerably inhibited like the death of trees caused by ultraviolet rays. By consequently, if the degree of ultraviolet radiation from the earth will increase due to the increase in the destruction of the ozonosphere, can achieve the conservation of forests planted by the man through the present invention. Using the method of plant trees of the present invention, forests can be formed planted by man endowed with lightning resistance ultraviolet.

Claims (15)

1. Group of plants formed by cultivation of seed plants whose chromosome sets are of type polyploid 2. Group of plants according to claim 1, in which the number of chromosomes is polyploid, such as triploides or tetraploides. 3. Group of plants according to claim 2, in which the seed plants are coniferous. 4. A group of plants according to claim 3, wherein the coniferous seed plants are selected from the group that includes Japanese cedar (in Latin, Cryptomeria japonica D. Don ) or Japanese cypress (in Latin, Chamaecyparis obtusa Endl .) . 5. Group of plants according to claim 4, in which the number of seed plant chromosomes is obtained by increasing the number of chromosomes by culturing plants comprising the step of artificially forming a vegetation considerably resistant to ultraviolet rays, as well as a mutation that occurs through the procedure of the colchicines and by their use in the mutation of the yolks. 6. Group of plants according to claim 1, in which the seed plants are coniferous. 7. A group of plants according to claim 1, wherein the coniferous seed plants are of Japanese cedar ( Cryptomeria japonica D. Don ) or Japanese cypress ( Chamaecyparis obtusa Endl .). 8. Method for planting trees, according to previous claims comprising the steps of selecting Seed plants of polyploid type and plant them. 9. Method for planting trees, according to claim 8, wherein the number of plant chromosomes Seed is polyploid, such as triploid or tetraploid. 10. Method for planting trees, according to claim 9, wherein the seed plants are of conifers 11. A method of planting trees according to claim 10, wherein the coniferous seed plants are Japanese cedar ( Cryptomeria japonica D. Don ) or Japanese cypress ( Chamaecyparis obtusa Endl .). 12. Method for planting trees, according to claim 11, wherein the number of plant chromosomes of seed is obtained by increasing the number of chromosomes by culture and mutation 13. Method for planting trees, according to claim 8, wherein the seed plants are of conifers 14. A method of planting trees according to claim 8, wherein the coniferous seed plants are Japanese cedar ( Cryptomeria japonica D. Don ) or Japanese cypress ( Chamaecyparis obtusa Endl .). 15. Plant cultivation method according to the preceding claims comprising the step of forming artificially a vegetation considerably resistant to ultraviolet rays compared to the vegetation constituted by triploid plants, by selective cultivation of plants whose number of chromosomes is greater than the basic number of the species.