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WO2015162626A2 - Dispositif d'échantillonnage à matrice trois voies pour feuilles et procédé associé - Google Patents

Dispositif d'échantillonnage à matrice trois voies pour feuilles et procédé associé Download PDF

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Publication number
WO2015162626A2
WO2015162626A2 PCT/IN2015/000177 IN2015000177W WO2015162626A2 WO 2015162626 A2 WO2015162626 A2 WO 2015162626A2 IN 2015000177 W IN2015000177 W IN 2015000177W WO 2015162626 A2 WO2015162626 A2 WO 2015162626A2
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WO
WIPO (PCT)
Prior art keywords
leaf
column
row
samples
leaf samples
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/IN2015/000177
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English (en)
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WO2015162626A3 (fr
Inventor
R. Dhandapani
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Indian Council of Agricultural Research
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Indian Council of Agricultural Research
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Publication date
Application filed by Indian Council of Agricultural Research filed Critical Indian Council of Agricultural Research
Publication of WO2015162626A2 publication Critical patent/WO2015162626A2/fr
Publication of WO2015162626A3 publication Critical patent/WO2015162626A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/0098—Plants or trees
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
    • G01N1/08—Devices for withdrawing samples in the solid state, e.g. by cutting involving an extracting tool, e.g. core bit
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/36—Embedding or analogous mounting of samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
    • G01N2001/2873—Cutting or cleaving

Definitions

  • the present invention relates to a sampl ing device. Particularly to a hand held device that faci litate in sampl ing leaves for D A based genetic purity testing of crop plants (varieties/ cultivars/ parental lines / hybrids) at seed production field and identify the contaminants if any. BACKGROUND ART
  • Seed qual ity has been held as sacred, being an important factor in the improvement of agriculture and agrarian societies.
  • the Rigveda, 2000 BC indicate the importance accorded to seed and the mother earth.
  • 5th century Kautilya Artha Shasthra, surapalas vrikshayurveda mentioned importance of seed and mentioned about seed treatments to ensure good germination.
  • Quality seeds are ones that are highly viable, genetically uniform and free from seed borne pathogen. Among various qualities of seeds like, germination, longevity etc., Genetic Purity (GP) of the seed lot is of top priority in order to have the fullest potential yield of the crop plants. Farm yield can be enhanced up to 10- 1 5 % by using genetically pure good quality seeds. The genuineness of the variety/hybrid is one of the most important characteristics of good qual ity seed. Maintenance of high level of genetic purity in hybrid seed lot is a major challenge to exploit the moderate level of heterosis. For instance, it is estimated that for every 1 percent impurity in rice hybrid seed, the yield reduction is 1 00 kg per hectare.
  • G P Genetic purity
  • GOT has many disadvantages, for e.g. it is time consuming (takes one full growing season for completion), space demanding and often does not allow the unequivocal identification of genotypes. Moreover, it is subjective, as several aspects of plant phenotype (morphology, yield, etc.) get influenced by environmental conditions. Moreover this testing approach, can detect only GP % and cannot be used to identify the contaminated seeds present in the seed lot and remove them to produce 100% genetically pure seed lot. Maintaining 100 % genetically pure crop stand at seed production field is very important, as it is a preventive approach.
  • Indian patent appl ication 2270/CHE/201 1 discloses DMA finger printing based genetic purity testing technique for commercial seed lot. The technique involves bulking of single leaf bits from 50 seedlings and analyzed through PCR involving SSR marker in 8 repl ications. Presence of extra alleles other than the allele 'specific' and 'unique' to the test variety is notified as off-type in one of the bulked sample of 50 seedlings. The extra allele confirms that at least one of the 50 seedlings tested is an off type.
  • Nas et al. (2000) proposed a two dimensional DNA sampling strategy involving a 10 x 10 grow out matrix for assessment of genetic purity of rice hybrid using A FLP molecular marker.
  • This strategy was original ly developed for hybrid rice seed production and it can be used for pure l ine varieties of rice and other crops.
  • 1 00 seeds were germinated in 10 row .x 10 column grow out matrix and two leaf bits from each seedling is collected for DNA based fingerprinting.
  • Each leaflet from respective row and column was separately bulked and used for identification of contamination with any other variety.
  • Rows and columns involving the off-type showed a fingerprint pattern different (with extra bands) from the control and were considered as impure rows and columns. Plants located on hi l ls where the impure rows and columns intersect were considered as suspected off-types.
  • US Patent US 8561346 B2 discloses an automated seed sampler including a sampling station; a sampler for removing material from a seed in the sampling station; a seed conveyer for conveying the seed from the sampling station to a compartment in a seed tray and a conveyor for conveying the material removed from the seed to a corresponding compartment in a sample tray.
  • the method of this invention comprises feeding seeds individually to a sampling station, removing a sample from the seed in the sampling station; conveying the sample to a compartment in a sample tray, and conveying the seed to a corresponding compartment in a seed tray.
  • the samples can be tested and the seeds can be sorted according to the results of the testing of their corresponding samples.
  • This patent teaches that each & every seed is sliced in to two halves and endosperm portion of seed is used for DNA fingerprinting. Other embryo portion is sorted and used for sowing, only if DNA based GP testing is qual ified or rejected. Though, this sampler collects and sorts the seed sample, but this sampler is si lent about bulking of seed sample that reduces the cost involved in DNA based GP testing.
  • molecular Marker Assisted Selection is the common methodology employed for developing new varieties/ hybrids in both public and private seed sectors in order to increase the efficiency of breed ing programme.
  • MAP Marker Assisted Seed Production
  • MAT Marker Assisted Seed Testing
  • Yet another object of the invention is to provide a method of forming the hand held device, which is capable of col lecting leaf samples from seed production field using three way sampling technique to identi fy the contam inants if any.
  • the afore-mentioned objectives are served by the present invention by way of providing a hand held device having a leaf sampler, matrix sorter, roll cartridge, tracker and counter.
  • the leaf sampler col lects three leaf samples from plants to be tested and del ivers them to matrix sorter.
  • the present invention also envisages that the matrix sorter segregates the leaves samples col lected from each plant in such a way so as to form a three way matrix. Also, the third sample of the leaf is retained separately by rol l cartridge for second time analysis, if required. Moreover, the present invention envisages that the tracker affixes a numbered label on surface of the plant leaf sampled, in order to reverse track contaminants from laboratory to seed production field and a counter to track the number of the leaf samples collected or to be collected.
  • the present invention also teaches a method of collecting, sorting, bulking and label ling leaf samples by placing a leaf from desired plant to be tested in a leaf holder of a leaf sampler for the purpose of collecting leaf samples; obtaining three disc shaped samples by punching the leaf using three punching rods of the leaf sampler; segregating two parts of said three leaf samples in removable containers arranged circularly in two circular row & column cassettes and retaining a third part of the said three leaf samples in a roll cartridge for further assessment to trace accurate contaminant, thereby forming a three-way matrix; and labell ing the leaf samples using a tracker for reverse tracking of the contaminant; and tracking the number of the leaf samples collected or to be collected for performing three way DNA sampling method.
  • Figure 2 I l lustrates left side, front perspective view of the hand held device.
  • Figure 3 I l lustrates exploded view of the hand held device.
  • Figure 4 I l lustrates cross sectional view of the hand held device.
  • FIG. 5 I l lustrates components of rol l cartridge and tracker in the hand held device.
  • FIG. 6 I l lustrates the components of counter in the hand held device.
  • Figu re 7 Diagrammatic representation of the hand held device.
  • Figu re 8 I l lustrates three-way matrix format of crop plants at seed production field.
  • Figure 9 I l lustrates three l ine (parents) multiplication system of hybrid rice seed production. DETAILED DESCRIPTION OF THE EMBODIMENTS
  • the present invention contemplates a hand held device (I) as in Figs. 1 & 2, to carry out matrix assisted multirex assay, particularly three way sampling method for assessing genetic purity of standing crop plants at seed production field.
  • the said device (1) simplifies the cumbersome three way sampling method by direct row and column bulking and helps in Marker Assisted seed Production (MAP).
  • MAP Marker Assisted seed Production
  • the three way sampling method involves collecting a large number of samples from a crop field. This sampling strategy also facilitates, DNA based genetic purity testing of standing crop plants and identify the contaminants.
  • This three-way sampling method involves collecting three leaf samples (round punches / leaf discs) from each plant and thereafter, two samples of leaf are arranged in a matrix while the third part is retained for further assessment. In matrix form, one part of every leaf is considered for row bulking, whereas the second part of the leaf is considered for column bulking. Thereafter, DNA is isolated from bulked rows and columns sample to reduce the cost of assay. The isolated DNA of respective column and row then undergo PCR assay with SSR markers to trace out the contaminant/ off type plant.
  • FIG. 1 & 2 illustrates perspective views of the hand held device, in accordance with an embodiment of the present invention.
  • the hand held device ( 1 ) would facil itate seed officials/ breeders / researchers to use leaf based three-dimensional DMA sampling strategy for testing genetic purity of standing crop plant and identi fy the contaminants during Marker Assisted seed Production (MAP).
  • MAP Marker Assisted seed Production
  • the hand held device ( I ) as in Figure 3, chiefi ly comprises five components - a leaf sampler (2), a matrix sorter (3), a roll cartridge (4), a tracker (5, 6) and a counter (7).
  • the leaf sampler (2) is used to col lect leaf samples (round punch of leaf disc) from standing crop plants in a field, which are to be tested.
  • the sampler (2) delivers the collected leaf samples to the matrix sorter (3) comprising of column (3a) & row cassettes (3b) in a desired orientation.
  • the matrix sorter (3) then, segregates the samples collected from each plant in such a way to form three way matrix.
  • Leaf sampler (2) in the hand held device ( 1 ) ( Figure 4) is used to punch out three small leaf discs from either single or three different leaves of each plant to be tested.
  • Leaf sampler (2) consists atleast of 3 components, i.e.
  • leaf holder ( 10) When a trigger (8) is pressed, leaf holder plate ( 10) moves downwards to position the leaf sample. The downward movement of whole leaf sampler is aided through punch guides ( 14 & 15) on both the sides and punching rods ( 1 1 , 12 & 13) in the middle.
  • the hand held device ( 1 ) (cross sectional view in Figure 4), when triggered using the trigger (8), col lects three leaf samples/discs from each plants to be tested. In an embodiment of the invention, one leaf from each plant is selected and inserted between leaf holder ( 10) and a bottom plate (9).
  • the leaf sample directly goes into the leaf sampler (2), for slicing the leaf in to three leaf discs which will be further considered for row and column bulking, and a further assessment of the probable contaminated sample, if required.
  • punching rods (1 1 , 12 & 1 3) punch three leaf discs in the leaf and mobilise them down to the matrix sorter (3) for segregation.
  • a cutting means ( 1 6) in the tracker, as wil l be explained later, slices glitter paper roll in to small label and the label is affixed/pinned on leaves of the plants being sampled.
  • the matrix sorter (3) col lects the three leaf discs of the leaf samples from leaf sampler (2).
  • the matrix sorter (3) comprises at least of three components; a removable column cassette (3a), row cassette (3b) and cassette cover (3c). Circularly arranged number of removable vials ( 1 8), corresponding to respective row and column samples, are present in both row and column cassettes (3a & 3b). The maximum number of vials denotes the matrix size of the bulking experiment.
  • Cassette covers (3c) with barcodes assists the hand held device ( 1 ) in protecting free fall and intermixing of sample once received inside row and column tube during post sampling handling in DNA fingerprinting laboratory.
  • the roll cartridge (4) in the hand held device ( 1 ) retains the third part of leaf samples for further, assessment of only probable contaminants for more accurate results (more clearly shown in figure 5).
  • the roll cartridge (4) comprises atleast of four components; tape holder (21 & 22), tap guider (23& 24), rol l cartridge holder (25), and main drive (26).
  • the tape holders (2 1 , 22) function in clockwise and anti-clockwise movements respectively to form a sandwich of tapes that contain leaf discs using tape guider (23). .
  • This rol led sandwich formed tapes enables tracing out contaminants accurately using third sample of all plant leaves tested for purity.
  • the added advantage of custom made sticky tapes are the presence of shallow wells and printed number, for quick removal of retained third leaf sample for DNA isolation.
  • the tracker (5 & 6) in the hand held device ( 1 ) enables labell ing of plants whi le sampl ing for genetic purity testing (figure 5):
  • the labelling of plants would help in tracking and identification of the contaminant plants that are identified using laboratory based MAM assay and remove them from seed production field to produce 1 00% genetically pure seeds.
  • the tracker comprises of atleast 4 components; a rol l holder (27), paper ejector (28 & 29), the cutting means ( 16), and an affixing means ( 1 7):
  • the roll of glitter paper is placed in roll holder (27) and guided through paper ejector (28 & 29) in such a way to protrude just above the leaves in leaf sampler and below the leaf holder.
  • the cutting means ( 1 6) and affixing means ( 17) present in the leaf sampler (2) assists in slicing the glitter roll in to small labels and labell ing the leaves of crop plants.
  • the counter (7) in the hand held device ( 1 ) enables the tracking of the number of samples col lected or to be collected ( Figure 6).
  • the counter (7) comprises atleast of three components; row sleeve (30), column sleeve (3 1 ) and indexing head (32).
  • the column sleeve (3 1 ) holds column cassette (3a) with the help of indexing head (32) and positions column tube to receive three way leaf samples.
  • the column cassette (3a) rotates and collects leaf samples when column sleeve (3 1 ) rotates.
  • the row cassette (3b) also rotates with the help of row sleeve (30) and indexing head (32) when column cassette (3 1 ) completes a full duty cycle (matrix size).
  • the encrypted numbers on row and column sleeves are used.
  • N number of plants
  • R row sleeve number corresponds to row tube number
  • M matrix size
  • C column sleeve number corresponds to column tube number.
  • Figure 8 illustrates three-way matrix sampling of 64 plants using the said hand held device (I), in accordance with an embodiment of the present invention.
  • each checker box numbered I to 64 denotes the individual plants with 3 or more leaves that are to be sampled using the device in 8X8 matrix format.
  • Example 1 For an instance, suppose 64 standing crop plants are to be tested using the said hand held device (I) in the seed production Held and arranged in a matrix of eight plants per row and eight plants in a column. Eight ready to use plastic vials are placed in to the cassette (3a) for column bulking and eight vials placed in to the cassette (3b) for row bulking ( Figure 7). As the device (1) is triggered, leaf disc from the first plant is pushed into first vial of the column cassette (3a) & row cassette (3b) from the leaf sampler (2). Thereafter, the column cassette (3a) rotates to. next vial to avail next plant sample. This is done in order to fill the sliced part of different leaf sample into the next vial.
  • each of the 8 vials is filled with one sliced part of different leaf samples.
  • row cassette (3b) does not rotate to aid in row bulking.
  • row cassette (3b) does not move in order to collect all 8 spliced parts of different leaf samples into one vial, which helps in forming first row bulking of the three-way matrix.
  • row cassette (3b) rotates to the next vial.
  • column cassette (3a) completes full duty cycle (eight rotational movements) and is back to its first vial with just one spliced part of a leaf sample already in it.
  • the row cassette (3b) fills all 8 different samples into one vial, and then rotates to the next vial.
  • This facilitates in making one complete row of the three-way matrix using the row cassette (3b) while fi ling only one part of each of the 8 columns in the three-way matrix, using column cassette (3a).
  • the matrix sorter (3) aids row and column bulking of the two spliced parts of the plant leaf sample.
  • This working of the hand held device (.1 ) to form an 8X8 matrix in above example is only exemplary to understand the working of the hand held device.
  • the said device enables NXN matrix depending upon the number of plants to be tested.
  • the hand held device ( 1 ) has the provision for collecting, sorting and labelling leaf samples from 400 standing crop plants, which is the minimum required number of plants examined by seed officer for assessing the genetic purity of a seed lot.
  • the collected leave samples of 400 standing crop plants are sorted both row and column wise yielding only 40 (20 each row and column) sample. These sorted ready to grind leaf samples used directly for DNA based genetic purity assessment of crop plants using microsatel lite SSR markers at DNA fingerprinting laboratory.
  • the collection of leaf samples is not limited to only 400 plants and can accommodate "n" number of plants leaf samples depending upon the vial used or the number of holes in the vials.
  • samples from 360000 standing crop plants were collected thus forming a 600X600 matrix.
  • samples from 1000000 standing crop plants were collected thereby forming a 1000X 1000 matrix
  • the invention describes an exemplary method of assembling this sampl ing device ( I ), which is capable to collect samples as per the three-way matrix sampling method.
  • the hand held device ( I ) being divided in to two halves, top and lower portion.
  • Top portion comprises leaf sampler (2) and its components.
  • Lower portion comprising matrix sorter (3), roll cartridge (4), tracker (5&6) and counter (7).
  • Leaf sampler (2) comprises at least of three components, i.e., leaf holder ( 10), three punching rods ( I I , 12, 1 3) and punch guiders ( 14, 1 5).
  • the whole set up of leaf sampler (2) is placed above the bottom plates (9) that contain three punching holes corresponding to the matrix sorter (3) and roll cartridge (4).
  • the elevated platform has been provided in-between the bottom plate (9) and leaf sampler (2) that creates the offset needed for insertion of leaf sample.
  • the punch guiders (14 ft 1 5) are fixedly placed on the main body of leaf sampler (2) that are attached with elevated platform.
  • the punch gu iders ( 14 & 1 ) are made of atleast three internally placed hollow pipes and a spring system.
  • the inner hol low pipe is attached with the leaf holder ( 10)
  • the outer is attached in the main body of leaf sampler (2)
  • the third hollow pipe referred here as middle pipe is attached to the trigger (8) and moves in-between outer & inner hallow pipes.
  • This arrangement guides the leaf holder ( 10), punching source and affixing means to enable precise leaf punches and move downward in to the tubes.
  • the spring is placed within inner most hallow pipe in order to assist initial quick movement of leaf holder ( 1 0) and ejection of trigger (8).
  • the leaf sampler (2) moves downward with the help of trigger pin that connects middle pipe of two punch guiders ( 14 ft 1 5) and three punching source (1 1 , 12, 13).
  • the easily removable trigger pin assists in replacing the components of leaf sampler.
  • Lower portion of the device ( 1 ) comprises of matrix sorter (3), roll cartridge (4), tracker (5 & 6), counter (7) and placed one after another serially in a horizontal perspective just below the punching and stapling sources.
  • the very crux and beauty of the device is its differential rotational access and its components.
  • the row sleeve (30) placed permanently in counter side indexing head (32) and helps in differential rotation of row cassette (3b).
  • the removable column sleeve (3 1 ) connects tracker (5 & 6) side indexing head (32) via row sleeve (30) and helps in continuous rotational movement of column cassette (3a), roll cartridge (4) and tracker (5 & 6).
  • the differential rotational access gear placed in-between row and column sleeve (30 & 3 1 ) helps in differential rotation of row and column cassette (3b & 3a).
  • tracker side plate consist roll cartridge and tracker components:
  • the drive initiated at column sleeve (3 1 ) rotates the column sleeve (3 1 ), rol l cartridge (4) and tracker (5 & 6) along with counter (7).
  • the invention also describes the method of col lecting leaf samples using the developed device ( 1 ).
  • FIG 4 i l lustrates a cross sectional view of hand held device ( 1 ). in accordance with an embodiment of the present invention.
  • the said device ( 1 ) collects leaf samples from crop plants.
  • three smal l leaf discs were punched by three punching " rods ( 1 1 , 12 & 13) present in the leaf sampler (2). Further, each of the three leaf discs pass through the corresponding punch holes ( 1 9 & 20), to move to the matrix sorter (3).
  • the matrix sorter (3) segregate all three discs of the leaf sampled in to row, column and retaining the third part separately, to form a three-way matrix for assessing genetic purity of standing crop plants in seed production field.
  • the row and column bulked leaf samples used for DNA isolation and further GP testing. This is done with the use of removable vials /tubes/containers (18) arranged circularly in two circular row & column cassettes (3a & 3b).
  • the two of the three holes. (19) in the leaf sampler (2) corresponds to row & column cassettes (3a & b). While the third hole (20) corresponds to roll cartridge (4) particularly, tape guider (24).
  • the circular row & column cassettes (3a &b) are capable to rotate from central axis namely row sleeve (30) column sleeve (3 I) respectively; hence, the vials (18) in the cassettes (3a &b) also rotate along with the respective cassettes.
  • the two parts of the leaf samples pushed through the holes (19) into the vials (18) Of the row & column cassettes (3a &b).
  • the cassettes (3a &b) rotate in order to fill the next vial (18) with the next leaf sample.
  • the matrix sorter (3) segregates two leaf discs of the leaf samples for row and column bulking, which is done by the rotating and positioning row and column cassettes (3a &b).
  • the circular cassettes (3a &b) facilitate row and column bulking.
  • One of the two cassettes facilitate for row bulking (3b) while the other for column bulking (3a).
  • FIG. 9 illustrates a three line (parents) multiplication system of hybrid seed production of rice, in accordance with an embodiment of the present invention.
  • the hand held device ( 1 ) has been used for detecting contaminant crop plants where three-line multiplication system exists during hybrid seed production.
  • three l ine (parents) multiplication system having A (Cytoplasmic Male Sterile (CMS) female), B (maintainer of A or A without steril ity) and R (male) line ( Figure 4) are used.
  • CMS Cytoplasmic Male Sterile
  • B maintainer of A or A without steril ity
  • R male line
  • 2 B lines are planted in adjacent to 6 A lines in order to multiply CMS female line during A * B crossing.
  • B l ine is harvested out first and later, the seeds produced by A line are harvested and used as female parent for hybrid seed production.
  • the device ( 1 ) can be effectively used in seed production field well before flowering of crop plants. In a similar way, the device ( 1 ) can also be effectively used for producing 1 00% genetically pure variety/hybrid seed of crop plants.
  • the present device being a hand held transparent device make it relatively easier to carry out error free leaf based three way sampling method for testing parental genetic purity of hybrid seed production field and identifying the contaminate, if any.
  • Row bulking and column bulking is automatic within in the matrix sorter (3) of the device (1) and ready to grind leaf samples can be directly used for DNA isolation.
  • the added advantage of this device (1) is presence of provisions for removing row and column cassettes (3b & 3a), which are loaded with (row & column wise) bulked leaf sample in a three way matrix format along with roll cartridge (4) contain all plant sample.
  • the roll cartridge (4) helps in retaining third part of the leaf samples and locating the plant from contaminated seed.
  • the device reduces the complexity of the method for assessing the genetic purity in a large number of samples. In addition, it also reduces cost and time. Further, the present device find its application in conjunction with leaf based three way sampling method in various central referral seed testing laboratory, state seed testing laboratory, DNA finger printing lab of public and private seed organisations for tracing the plant originated from contaminated seed using molecular markers and identify the contaminants, if any. Along with three way sampling method the present device can also be effectively used at fields to identify the parental line purity of hybrid seeds at various public seed testing laboratories (STL ' s)/ seed companies/SAU's/ research institutes in India and Globally

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Abstract

La présente invention concerne un dispositif manuel simple et convivial (1) pour la collecte, le tri, le regroupement et le marquage d'échantillons de feuilles en vue d'évaluer la pureté génétique de plantes cultivées, comprenant un dispositif d'échantillonnage de feuilles (2) pour collecter des échantillons (disques) de feuilles à partir de plantes souhaitées à tester ; un trieur à matrice (3) couplé au dispositif d'échantillonnage de feuilles (2), le trieur à matrice (3) étant conçu pour séparer en deux parties lesdits échantillon de feuille dans un format de matrice trois voies pour un regroupement de type rangée et colonne ; une cartouche à rouleau (4) pour stocker et garder une troisième partie de l'échantillon de feuille séparé pour une évaluation ultérieure visant à suivre un contaminants précis ; un dispositif de suivi (5, 6), fonctionnellement couplé à au dispositif d'échantillonnage de feuille (2), pour marquer les échantillons de feuille en vue du traçage inverse de la plante contaminante ; et un compteur (7) pour suivre le nombre d'échantillons de feuille échantillons collectés ou à collecter pour mettre en œuvre le procédé d'échantillonnage d'ADN trois voies.
PCT/IN2015/000177 2014-04-21 2015-04-21 Dispositif d'échantillonnage à matrice trois voies pour feuilles et procédé associé Ceased WO2015162626A2 (fr)

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IN180/DEL/2014 2014-04-21
IN180DE2014 2014-04-21

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WO2015162626A3 WO2015162626A3 (fr) 2016-01-21

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108801673A (zh) * 2018-06-04 2018-11-13 华中农业大学 一种便携式多功能植物叶片打孔器
CN114858505A (zh) * 2022-07-07 2022-08-05 中国农业科学院蔬菜花卉研究所 植物叶片采样仪
CN115105129A (zh) * 2022-07-05 2022-09-27 北京积水潭医院 一种自动化核酸采样检测试管收集装置
CN115235806A (zh) * 2021-04-23 2022-10-25 河南科技学院 一种适于木香薷叶表皮腺毛观察用的取样器

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108801673A (zh) * 2018-06-04 2018-11-13 华中农业大学 一种便携式多功能植物叶片打孔器
CN115235806A (zh) * 2021-04-23 2022-10-25 河南科技学院 一种适于木香薷叶表皮腺毛观察用的取样器
CN115105129A (zh) * 2022-07-05 2022-09-27 北京积水潭医院 一种自动化核酸采样检测试管收集装置
CN115105129B (zh) * 2022-07-05 2023-06-02 北京积水潭医院 一种自动化核酸采样检测试管收集装置
CN114858505A (zh) * 2022-07-07 2022-08-05 中国农业科学院蔬菜花卉研究所 植物叶片采样仪
CN114858505B (zh) * 2022-07-07 2022-09-30 中国农业科学院蔬菜花卉研究所 植物叶片采样仪

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