s.

GERMINATION TESTING
IJ.AS'rVAl1/A/AI1, /Hi. GUPTi\, RAMEGOW/J/\ & M.V REDDY
Germination testing is considered as the most important quality ;est in evaluating the planting value ofa seed lot. The ability of seeds to produce normal seedling~ and plants later on is measured in terms of germination test. Testing of seeds under fieldconditions is normally unsatisf~lctory as the results cannot be repnxJuced with reliability. Laboratory methods then have been conceived wherein the external factors are controlled to give the most uniform. rapid and complete germination. Testingeonditions in the labomtory have been standerdized toenablc the test results to be reproduced within limits :'l"nearly as possible:.L<;those determined by random sample variations. 5.1 O~jective The ultimate objective of seed germination testing is to obtain information with rc.<.;pect to the planting value of the seed and to provide resulL" which could be used to compare the value of different seed lots. 5.2 Definition

Germinationof a seed lot in a laboratory is the emergence and development of the seedling to a stage where the aspect of its essential structures indicates whetheror not it is able to develope further into a satisfactory plant under favourableconditions in soil (fSTA. 1985). "These essential structres are a well-developedand intact root system. hypocotyl. plumule and one or two cotyledons according to the specic.<;. Seedlings cannot be evaluated ina germination test until these essential structures are clearly identifiable and the reported percentagegermination expresses the proportion of seed,>which have produced normalseedlings within the period specified for each species. 5.3General Principles Germinationtests shall be made with seeds from the pure seed fraction of a
purity test. A . . minimum of four hundred seeds are required in four replicates of 100

45

seeds each or eight replicates of 50 seeds each or 16 replications of 25 seeds each depending on the size of seeds and size of containers of substrate.

TI1e seeds shall receive no pretreatments

excepting those recommended

in

Table 5.1. If additional tests are undertaken after any other pretreatment, the result and pretn:,ltnlent must be reported under other determinations on the certificate. The seeds. ,irranged in replicates. ,Ire tested under favourable moisture conditions and in accordance with the metl1<xlsprescribed in Tahle 5.1. After the period indicated in "I "able 5.1 the replicates ,Ire examined and counts made of the seedlings and seeds in various categories required for reporting as presented elsewhere. 5.4 General Requirements ti!r Germination

Seeds require certain condit ions for normal germ in,ll!on. The most important requirements arc substrata. moisture. tempcr;!lurc ;lJ1dlight.
5.4.1. Suitable ".I"tratum

The substrata serve as a moisture reservoir and provide a surface or medium for which the seeds can germinate and the seedlings grow. "111e commonly used substrates are paper. sand and soil.
5.4.I.a Paper substrate

Most widely l\I>edpaper substrates are filter paper, blotter or towel (kraft paper). these arc easy to handle. versatile and compar,ltivcly cheap. Specifications for germination paper arc given in Appendix. 5.4.1.bSand The specification for sand substrata is given in Appendix. It may be necessary to wash and sterilisc the sand before use. For reuse of sand it must be washed, dried and resterilized. Sand which has been used for testing chemically treated samples. should preferably be discarded without being reused. if however. it is reused it should be ascertained that chemicals which may have accumulated in the sand do not cause phytoxic symptoms.
5.4.1.<'Soil

Soil should be or good quality. non-caking and free from any large particles. It must he reasonably free from weed seeds, bacteria. fungi. nematode~ or toxic substances. which might interfere with the germinmion of seeds. the growth of 46

seedlings or their evaluation. Soil should allow adequate aeration for germ ination when water is added with a pI I of 6.0- 7.5. Bcf(}re use soil may require sterilisation and soil is not recommended for reLL'ie.
5.4.2. Adequak ;\<Ioi~ture or Water

High concentration of water at cellular level is necessary fix the seed to start germination. Mobilisation of food requires hydrolysis (hreaking down process) to transport materials from storage to growing tissues. Moisture is supplied to the seeds through the substratum. (ienerally. the moistened substrata is sufricient to rehydrate to 10-80 per cent. However. the moment the rediclc emerges. additional moisture contributes better seedling growth. In the case of vegetahle seeds. care is necessary in moistening the substrata. Too much water would allow fungal growth and decay of sceds. The general specifications for water are: It should he free from organic or

inorganic impurities. 'lhe pi I value should he within the range of ().() to 7.5. If the usual water supply in the lahoratory is not satisfactory. distilled. de-ionised water may be used. To ensure the quality or water being w..ed. an analysis should he obtained from time to time.
5.4.3. Fa\'oUl'"dble temperature

Germination occurs under different ranges of temperatures provided the sced is given adequate moisture. Temperature is not as critical as water requirement during the test. Seeds of most of agricultural and horticultural crops germinate in the temperature range of 1OC-15('. Some seeds germinate oeHer at constant temperature. Others require an alternating temperature. Temperature control docs provide the comparable conditions exactly under which test can he repeated. Temperature control is also necessary to overcome dormancy wherever it of

occurs. Exposure of seeds to the temperature at 40C or higher. alternation

temperature. low temperature applications are the easiest and safest method to overcome seed dormancy although methods to overcome dormancy by chemic..'11 treatments do exist.

Therefore. the temperatures prescribed in Table 5.1 should be determined at the level of the seeds on the substrate.
5..U. Light

There are crops for which light is not required during germination test. 47

However. presence of light is desirable to enable the evaluation of seedlings easier and with greater certainty. Other crops like lettuce and tobacco require light during germination on the test.

5.5 Equipments for Germination Test

The equipment and materials that are required to conduct the germination test are given in Appendix I.

5.6 Procedures
5,(>.1. Working sample

Four hundred seeds are counted at random from the well-mixed pure seed. RepliC<ltesof 100 seeds are normally used. spaced sufficiently far apart on the seed bed to minimise tne effect or adjacent seeds on seedling development. To ensure adequate spacing. split replicates of 50 or even 25 seeds may be necessary. particularly where there is seed-borne disease. When the seeds are heavily infected it may also he necessary with a paper suhstrate to change the substrate at an intermediate count. Testing f()ur hundred seeds is recommended on seed law enforcement. seed certification and service samples.
5.6.2. Methods usin~ paper

Paper substrates arc used for the following methods: (a) TP (Top of Paper) : ;\s the name indicates. the seeds are placed directly on one or more layers of moist filter or blotter papers in petridishes. These petridishes are tightly covered with lid and placed inside the germination cabinet. The relative humidity in the cabinet must then be maintained to

95-99% to prevent drying out during Lestperiod.

(b) BP (Between Paper) : The secds are germinated between two layers of paper. This may be achieved by loosely covering the seeds with an additional layer of paper or by placing the seeds in rolled towels. The rolled towels are to be placed inside the germ inator in an upright ~ition. (c) PI' (Pleated Paper) : Seeds are placed in pleated strips. The paper may have 5-10 pleats which can be made in the laboratory. Each pleat may 48

have LO seeds.TQepleatedstrips are kept in moistenedbreadboxesto

ensure uniform moisture conditions. This method may be used as an alternative where TP or BP are prescribed.

5.6.3. Methods using sand

Sand is used as follows: (a) TS (Top of Sand) : The seeds are pressed into the surface of the sand.

(b) S (In Sand) : The seeds are planted on a levelled layer of moist sand and covered with 10-20 mm of uncompressed sand depending on the size of the seed. To ensure good aeration it is recommended that the bottom layer of sand be loosened by raking before sowing. Sand may be used instead of paper. even if not prescribed in Table.5.l. when theevaluationof a diseased sample proves impracticable because of the contaminationof the paper substrate.
5.6.4. Methods using soil

Since it is generally difficult to obtain consistency in soil or artificial com post.

it is not recommended as a primary testing substrate.

5.6.5. Mofiture and aeration

The substrate must all times contain sufficient moisture to meet the requirements for germination. However. moisture content must not be excessive. or aerationmay be limited. The initial quantity of water to be added will depend on the natureand dimensions of the substrate and also on the size and species of the
seed to be tested.

The amount of water to be added to the sand can be calculated as follows (Anonymous,1952). mlofwater to be addedto each 100 gramof sand

118.3 ml of sand Wl. of 118.3 ml of sand in gm

(20.2--8.0)

The amount of water provided by this formula is satisfactory for seeds of the size of mustard. for larger seeds slightly more and for smaller seeds slightly less

49

water should be added.

Subscquentwatering

should be avoided wherever possible as it is likely to

increase the variability between replicate..~and between tests. Therefore, precautions should be taken to ensure that the substrate may not dry out and that sufficient water is supplied continuously during the test period. Special measures for aeration arc not usually necessary for TP and HP tests. For OJ>,However, care should be taken that rolled towels arc loose enough to allow sufficient air around the seeds. ror the same reason the material covering the seeds in sand or soil tests should not be compressed.
5.6.6. Temperature

Temperatures are prescribed in Table 5.1 and should be determined at the level of seeds on the substrate.They should be as uniform as possible throughout the germination apparatus and care should be taken that the temperature of tests does not exceed the level prescribed and variation due to the apparatus should not be more than ~ 1c.

Where alternating' temperatures are indicated, the lower temperature should usually be maintained for 16 hours and the higher for 8 hours. If alternation of temperatures cannot be controlled over week-ends or public holidays, the test should be kept at lower temperature.
5.6.7.Light Seeds of most of the species in Table 5.1 will germinate either in light or in darkness. However. illumination of the substrate from artificial source or by day light is generally recommended for better seedling development to avoid etiolation and also to detect seedlings having chlorophyll deficiency.

Specific recommendations for light or darkness. respectively. are given in the additional directions column of TableS.L 5.7 Choice of Method When alternate methods indicated in Table 5.1 one of them (any combination of substrate and temperature) must be used. The choice of method will depend largely on the facilities and the experience of the Seed Testing Laboratories.

50

5.8. Pretreatments for Germination For various reasons (e.g. physiological dormancy. hard secdedness. inhibitory substances) a considerable number of hard or fresh seeds may remain at the end of the germination test. In order to prevent these non-germination and to have complete germination various kinds of pretreatment" arc recommended and given in Table 5.1. These pretreatments include dry storage. prechilling (treat the moist seeds at a temperature of 5 -1DoC for about seven days) preheating (at 30 -35( 'j. light (750-1250 lux from cool white lamps for 8 hrs per day). potassium nitrate (0.2% KNO3). gibherellic acid (GA3 0.0)-.1 %) application etc. . For hard seeds. puncturing the seed with a needle away from embryo. mechanical sC<lrification and acid scarification are recommended. Similarly for removing inhibitory substances. prewash the seeds by running water at a temperature of 2:'i(' and bring back these prewashed secds to its original moisture content. 5.9 Dumtion of the Test Duration of the test for individual species is indicated in Table :'i.1. The duration of the treatment required to break dormancy before or during the test is not taken as part of the germination test period.

The time of first count is approximate. but must be sufficient to permit the seedlings to reach a stage of development which allows for accurate evaluation. The time indicated in Table 5.1 refer to the highest temperatures. If lower temperature is choosen. the first count may have to be postponed. The tests lasting 7-10 days. intermediate count" to remove seedlings which are sufficiently well developed are recommended in order to make counting easier to prevent them from affecting the development of other seedlings. The first count may be omitted. if the first test is conducted in sand. If the maximum germination of the sample has been obtained before the end of the prescribed test period. a test may be terminated. The seed testing laboratory on request of producer may release the result of seed germination on the basis of first count if the sample in question meets the minimun limits of germination for certification/labelling. 5.10 Seedling Evaluation Seedings which have reached a stage when all essential structures can be 51

accurately assessed. shall be removed from the test at the first or any other intermediatecounts. Badly decayed seedlings should be removed in order to reduce the risk of secondary infection. but abnormal seedlings with other defects should be left on the subtrate until the final count.
5.10.1 Categories 5.10.l.a. of seedlings

Normal seedlings

Normal seedling is one which shows the capacity for continued development into mature plant when grown in good quality soil and under favourable conditions of water supply. temperature and light. lhis capacity for continued development depends upon the soundness and correct functioning of the developing structures (Fig. 5.1 and 5.2) during germination. According to the International Seed Testing A<;sociation (1985). seedlings to be classified categories : as normal seedling. must conform with one of the following

(a) Intact seedlings: Seedlings with all their essential developed complete in all proportion and healthy.

structures. well

(b) Seedlings with slight deJects: Seedlings showing certain slight defecL<;of their essential structures provided they show an otherwise satisfactory and balanced development comparable to that of intact seedlings of the same test.

(c) Seedlings with secondary inJections: Seedlings which are seriously infected by fungi or bacteria are classified as normal. if it is evident that the parent seed is not the source of infection. and if it can be determ ined that all the essential structures were present.
5.10.l.b. Abnormal seedlings

An abnormal seedling is one which does not have the capacity to develop into a normal plant when grown in the soil under favourable conditions because one or more of the essential structures is irreparably defc<.:tive.

Three major classes of abnormal seedlings arc:

(a) Da11lagedSeedlings: Seedlings with any of the essential structures missing or so badly damaged that balanced development does not OC<..llr. lbe damage to the embryo in the seed usually results from external cause i.e. mechanical handling.

52

heat, drought or insect damage which leads to the abnormalities. (b) Deformed or unbalanced seedlings: Seedlings with weak and unbalanced development which may be caused by internal disturbances of physiologicalbiochemical character. Such internal disturbances, however, are often due to the earlier external disturbances such as unfavourable growing conditions of the parent plants, poor ripening conditions for the seed, premature harvesting, effect of herbicides or pesticides and inappropriate storage conditions or ageing of the seed. (c) Decayed seedlings: Seedlings with any of their essential structures s6 diseased or decayed as a result of primary infection that normal development is prevented. These may result from the external or internal seed borne diseases.
5.10.l.c. "lult~crm ~ced unih

Seeds which arc capable of producing more than one seedling. Several types of seed units can produce more than one seedling e.g. unseparated schizorcarps of umbelliferae, clustersorlJcta vulgaris, fruiL'; of tectona grandis, polyembryonic seeds. In such cases only one normal seedling is counted for determining the germination percentage.
5.10.1.d. Ungcnninated ~eed

Seeds which have not germ inated by the end of test period when tested under the conditions prescribed in Table ).1 arc classified as follows:
I

(a) Hard seeds: Seeds which do not absorb moisture till the end of the test period and remain hard. (b) Fresh seeds: Seeds which are neither hard nor have germinated but remain clean and firm and apparently viable at the end of the test period. The viability of the fresh seeds may be determined by tetrazolium test.

(c) Dead seeds: Seeds at the end of the test period arc neither hard nor fresh nor have produced any part of a seedling. Often dead seed collapses and a milky pastecomes out when pressed at the end of the test.
5.10.2 Seedling descriptions

As per the ISTA Rules (1985 Para 5.2A.A. and 5.l.5.A. the following is the detaileddescription for normal and abnormal seedlings. These have to be taken intoaccount while evaluating the seedlings.
53

Detailed descriptions for normal and abnormal seedlings of representative genera of some species are illustrated in line diagrams (Fig. 5.3. to 5.14). Like wise Photographs depicting normal and abnormal seedlings in respect of some species may be seen from Page No. 86 to 92.
5.10.2.a. Normal seedlings

1. Intact seedling (Fig. 5.1 & 5.2) Depending on the species being tested, a combination ofsome of the following essential structures are generally found: a well developed root system having a long and slender primary root, usually covered with numerous root hairs and ending in a fine tip.

secondary rooL,;or seminal roots instead of one primary root in certain genera, includingAvcna, Hordeum, Secale, Triticum, Triticosecale. a well developed shoot axis consists of a straight and usually slender
elongated hypocotyl in seedlings showing epigeal germination.

a well developed epicotyl in seedlings showing hypogeal germination. both an elongated hypocotyl and epicotyl in some genera with epigeal germination. an elongated mesocotyl in certain genera of the Gramineac. a specific number of cotyledons, i.e., one cotyledon in monocotyledons or exceptionally indicotyledons (it maybe grecnand leaf-like or modified and remain wholly or partly within the seed). two cotyledons in dicotyledons (in species with epigeal germination they are green and leaf-like). In species with hypogeal germination they are hemispherical, fleshy and remain within seed coat. green expanding primary leaves. a terminal bud or shoot apex. a well developed straight coleoptile in Gra11lineae containing a green leaf. 54

2. Seedlingswith the following slight defect Primaryroot with limited damage of slight growth retardation, defective but withsufficientwell developed secondary roots in specific genera ofLequminosae (e.g. Phaseo/us, Pisum, Vicia); Gramineae (e.g. Zen); in all genera of Cucurhitaceae(e.g. Cucumis, Cucurhita, Citrullus) and Ma/vaceae (e.g. Gossypium). only two seminal roots in A vena, Hordeum, SeGGIe,Triticum, Triticosecole. hypocotyl, epicotyl or mcsocotyl with limited damage cotyledons with limited damage only one normal cotyledon in dicotyledons Primary leaves/leafwith limited damage and healthy termianl bud Colcoptile with limited damage, loosely twisted forming a loop,coleoptile with a green leaf not extending to the tip but leading at least half-way upto the colcoptile.

5.IO.2.b. Abnormal seedings

One or a combination of the following defects in the seeding renders it abnormal.

I.

Primary root: 1. stunted 2. stubby 3. retarded 4. missing 5. broken 6. split from thetip

55

7. .constricted

8. spindly
9. trapped in the seed coat 10. with negative geotropism 11. glassy 12. decalyed as a result of primary infection 13. only one seminal root or none. Note: Secondary roots or seminal roots showing one or more of the above defect" are abnormal and cannot replace an abnormal primary root in cases where the presence of several secondary roots (e.g. Cilcumis) or at least two seminal roots (e.g. Triticum) determine the value of a seedling. II.
The hypocotyl. the epicotyl. the mcsocotyl:

1. short and thick 2. deeply cracked or broken 3. split right through 4. missing 5. constricted 6. tightly twisted 7. bent over 8. forming a loop or spiral 9. spindly 10. glassy

56

11. decayed as a result of primary infection Ill. The cotyledons (apply 50% rule) : 1. swollen or curled 2. deformed 3. broken or otherwise damaged 4. separate or missing 5. discolourcd 6. necrotic 7. glassy 8. decayed as a result of primary infection
Note: Damage or decay of the cotyledons at the point of attachment to the seedling axis or adjacent to the shoot apex renders a seedling abnormal, irrespective of the 50% rule. Special cotyledon defects for Allium species. g. short and thick 10. constricted

11. bent over 12. forming a loop or spiral 13. without a definite 'knee' 14. spindly IV. The primary leaves (apply 50% rult?): 1. deformed 2. damaged 57

3. missing 4. discoloured
5. necrotic
'.

6. decayed as a resull of primary infection 7. normal shape hut less than 114normal size.

V.

The terminal bud and surrounding tissues: I. Deformed 2. damaged 3. missing

4. decayed as a rc..<.;ult of primary infection

Note: If the tenninal hud is defective or missing. the seedHng is abnonnal. even when one or two axillary huds (c.g. Phaseolus) or sh(x)ts(c.g. Pisum) have developed. VI. The coleoptile and first leaf (Gramineae) : The colcoptile : 1. defonned 2. damaged 3. missing 4. with the tip damaged or missing 5. strongly bent over
'It,.

6. forming a loop or spiral

7. tightly twisted 8. split for more than one-third of the length from the tip 58

9. split at the base 10. spindly II. decayedas a result of primary infection The first leaf: 12. extending less than halfway urto the colcop!ile U. missing
14. shn~dded or otherwise deformed.

VII.

The seedling as a whole: 1. deformed 2. fractured 3. cotyledons emerging before the root 4. two-fusedtogether 5. pe~isting endosperm collar 6. yellowor white 7. spindly
8; glassy

9. decayedas a result of primary infection.

50% rule: Seedlings are considered as normal if half or more of the total coty lcdon tissue/primary leaf is functional but abnormal when mme than half of the cotyledon tissue/primary leaf is not functional and defective.
5.11 Retesting

If the results of a test are considered unsatisfactory it shall not be reported and a second test shall be made by the same method or by alternative method under the 59

1!
following circumstances.

RepliC<ltes perlormance

is out of tolerance. Results being inaccurate due to

wrong evaluating of seedlings or counting or errors in test conditions. Donnancy persistence or phytotoxicity or spread of fungi or bacteria.

5.12 Reporting Results

The result of the germ ination test is calculated as the averages of 4 x 100 seed replicates. 1\ is expressed as percentage by number of normal seedlings. lhe percentage is calculated to the nearest whole number. The percentage of abnormal seedlings. hard. fresh and dead seeds is c<llculated in the same way. These should be entered on the Analysis Certificate under appropriate space. If the result is nil for any of these categories it shall be reported as '0' instead of leaving the appropriate column blank. 5.13 Use of Tolenmces For the use of tolerances. appropriate table which is given in flow chart 12.1 in Chapter 12.should be used.

60

LI

Table 5.1 Germination Methods

Prescription for

Addtional directions including recommenda-

Crop

Botanical name

Substrata

Temp. (C) 4

First count (days) 5

. Final count (days) 6

tion for breaking dormancy

FIELD CROPS aI .... CEREALS Barely Paudy Triticale Wheat MILLETS Barnyard milIet Commonmillet Echillocloa trumelltacea Pallicum miliaceum TP TP;BP 20-30 20-30;25 4 3 10 7
Prcchill,

Hordeum vulgare Olyza sativa Triticosecale wiltmack Triticum spp.

BP;S' BP;TP;S UP TP;BP;S

20 20-30;25 15-20 20

4 5 4

7 14 7 R

Preheat(30-35C), prechill, Cii\.~

Preheat (5()OC) soak in w,iler or

IINO:l24 hrs.

GI\:I, Prcchill Preheat

KNO:l, C;A~

(Prasa millet,
Hogmillet)

1 Finger millet Italian millet (Foxtail millet) Kodo millet Little millet Maize Pearlmillet
0) I\)

2 Eleusille eoraealla Setaria italica Paspalum scorbieulatum Panicum miliare and P. sumatrense lea mays Penllisetum typhoides Sorghum bicolor Cieer arietillum Vigna mullgo Lathyrus satil'us Vida sativa Vigna unguiculata PhaseDIus vulgaris Vigna radiata Do/ichos biflol'u.\'

3 TP; BP TP;BP TP TP;BP BP;S TP;BP TP;BP BP;S BP; S S;BP BP;S BP; S BP; S BP;S BP

4 20-30 20-30 20-30 20-30 20-30;25 20-30 20-30;25 20-30;20 20-30;2..'3 20 20

20-30; 2..'3

5 4 4 7 4 4 3 4 5 4 5 5 .5 5 5 3

6 8 10 20 7 7 7 10 8 7 14 14 8 9 8 5 Prechill KNO3

7 0.2% KNO3 (2-3 hrs)

0.2% KNO3 (2-3 Ius) Prechill

(Bajea) Sorghum

PULES
Bengalgram :Blackgram Chickling vetch Common vetch Cowpea French bean (Rajmash) Greengram Horsegram (Kuthi) 20-30;25 30

20-30;25;20

--

----

----2 3 S;BP BI'; S BP; S BP; S UP; S 4 20;20-30 20-30;25 20 20 30 5 4 5 5 5 4 6 10 10 10 8 6 I'rechill 7

Indianbean(Scm)
Kidney bean

E.ablabpurpureus Viglla acolli/ifolia Lells cuEillaris Pisum sa/il'Um Cajallus cajmr

(Moth bean) Lentil Peas Pigeon pea (Redgram) OILSEEDS Castor

0) (,)

Ricillus commullis Arachis hypogea l.illillum usi/a/issimum Brassica jullcea Brasslca lIigra Guizo/ia abyssillica Brassica lIapus Ertlca satim Carthamus thle/orius Sesamum indieum Glycine max
Hetiall/hus (IIIIIUUS

AI'; S BP; S TP; BP TP TP TP TP TP;BP II'; BP;S TP UP; S AP;S

20-30 20-30;25 20--30;20 20--30;20 20--30;20 20-30 20--30; 20 20 20--30; 25 20--30 20--30; 25 20-30-25; 25

7 5 3 5 5

14 10 7 7 10 14 7 7 14 6 8 10 Ethrel (25 ppm) 48 hrs Remove shells, Preheat -4!tC Prechill Prechill, KNO.3 Prechill, KNO3 Prcchill

'Groundnllt Linseed Mustard Mustard (Alack) Niger (Ramtil) Rape Rocket salad (Traramira) Safflower Sesamum (TiI) Soybean Sunflower

5 4 4 3 5 4

2 FIBRE CROPS Cotton Jute (PalSan) Roselle (mesta) Sunnhemp FORAGE CROPS Bird wood grass (Dhaman) Blue panic en
"'"

:1

Gos.\ypium spp. CorcllOrusspp. Hibiscus spp. Croto/ariajullcea Cellchrus setigerus Pallicum alllidota/e Cellchrus cilliaris CYllodolldacty/oll Cyamopsis tetragall%bus Alldropogall molltallus Pallllisetum pedicellatum Trifolium a/exalldrium Trigollella foellum-graecum

HI'; S TP; 131' HI'; S HP;S TP TP TP;S TP BP TP TP TP;BP TP;BP

20. :10:25 :10 20-30 20-30 20-35 20-30 20-35

20-30;30

4 .3 4 4 3 7 7 7 5 7 7 3 5
"

12 5 H 10 14 28 28 21
14

Hot water(H5OC-lminute)

Preheat (40()q

Preheat; Prechill. KNO3 Prechill. KN03; Light

I;3uffel grass Burmuda grass (Doob) Cluster bean

20-35 20-30 2-35 35;20-35 20

Dharaf grass Dinanath grass Egyptian clover (Berseem) Fenugreek (Methi)

28 28 7 14

Prechill at S()Cfor two weeks H2 S04 5% for 5 min.

20-30;20

1 Guinea grass Indian clover (Senji) Johnson grass Lucerne Marvel grass Napier grass Oat Para grass Rice bean (Red bean) Rye Rye grass Sataria grass (Nandi grass) Shaftal Style Sudan grass

2 Pallicum may/mum Meli/olus illdica Sorghum halapellse Medicago sativa

Dichanthium all1lUlatum

3 TP TP;BP TP;BP TP;BP TP BP BP; S TP BP TP; BP; S TP TP TP;BP TP TP;BP

4 15- 35; 20-30 20 20-25; 20-30 20 20-30 20--30 20 20-30 20-30 20 20-30; 15-2'5; 20 20-35 20 20-35 20-30

5 10 4 7 4 7 5 5 4 5 7

6 28 7 35 10 21 10 10 21 8 7 14 21 7 Prechill KNO3

7
Prechill, KNO.3

Prechill

C7I
01

Pe/!/Iisetumpwpureum A vella sativa Braclliaria mutica Viglla umbel/ala Secale cereale Lotium pare/l/Ie Setaria anceps Trifolium resupinatum Stylosanthus spp. Sorghum sudallellSe

Preheat 30-35OC,Prechill,

Prechill; GA1 Prechill, KNO3 KNO3

4 4

10 10

H2SO4 Prechill

2 Teosinte Velvet bean Euehlaena mexic(lIIa Stizelobium spp,

3 HI'; S BP;TP

4 20-30; 25 0-30

6 7

7 GAl \000 ppm - 24 Ill's.

14

GREEN MANURE AND MISCELlANEOUS CROPS Dhainch Hemp Indigo

Sesbania spp, Cannabis sati!'a Indigofera hirsuta

TP;BP TP;BP HI'

20--30 20-30; 20 20-30 -

5 3

7 7 14

Rub seed coat on Sand papcr. Continue test for a further 5 days if hard seeds have begun to imbibc,

0)
0)

Buck wheat Caraway Chicory Cumin Garden cress Lotus Poppy (Opium) Purslane Sugarbeet Tobacco Water cress

Fagopyrum eseulentum Carum car!'i Cichorium intybus CumoU/mcyminum Lepidium sa/ilIum Lotus comiculatum Papal'e,' somniferum Portulaca oleracea Beta I'ulgaris Nicotimla tabacum Nastutricum officiI/ale

TP;BP TP TP TP TP TP;BP TP; TP;BP TP; HI'; S TP TP;BP

20-30; 20 20-30 20-30; 20 20-30 20-30; 20 20-30; 20 20 20-30 20-30; 15-25 20-30 20-30

4 7 5 5 4 4 5 5 4 7 4

7 21 14 14 10 12 to 14 14 16 14 Prechill Prechill Prechill Prechill Prewash multigerm 2 hrs; monogerl11 4 hrs, KNO3 KNO3

---1 2 3

.."....

4 5 6 7

- - .

VEGETABLES CUCURBITS Ashgourd Bittergourd Bottlegourd Cucumber Indian squash Little gourd Longmelon & Muskmelon Pointed gourd Bellillcase hispida Memo,.dicachamllti Legella,.iaSlcem,.w Cucumis satil'us Praecit,.ullus fistulosus
(j)
"'-I

S BP; S HP; S
. TP;BP;S

30-35 20-30;30 20-30 20-30;25

.5 4 4 4

14 14 14 8

Light

Coccillia gmlldis Cucumis melD T,.icllOsallthus dioica Cucu,.bita maschata

BP; S S

20-30;25 30-35

8 14 Dark, GA3 500 ppm 24 hrs, Remove seed coat.

Pumpkin (Khashiphal) Ridge gourd Snake gourd Snap melon Sponge gourd

BP; S BP; S S BP; S BP; S

20-30;25 30 30-35 20-30;25 20-30;30

4 4

8 14 14 Dark, GA.3500 ppm 24 hrs. Remove seed coat.

Luffa acutallgula T,.icllOSallthus allguilla Cucumis melD Luffa cylilld,.ical Luffa aegyptiaca

4 4

8 14

2 Summer squash Water melon Cucurbita pepo Citrullus /al/atus

3 BP; S BP;S BP; S TP;BP TP;BP BP; S BP; S TP;BP

4 20-30; 25 20-30;25 20-30; 25 20-30 20-30 20-30 20-30;20 20-30

5 4 5 4 7 7 4 5 5

6 8 14 8 14 14 21 10 14 KNO3 KNO3

Cucurbita maxima Winter squash FRUIT VEGETABLES Brinjal Chilli Okra Rat tail (Mungra) Radish Tomato So/al/um me/ol/gel/a Capcicum spp. Abe/moschus escu/el/tus Rapha1luscaudatus Lycopercicum escu/entum Allium pOl.rum Dioscoria spp. Allium cepa So/al/um tuberosum
0'1 OJ

BULB AND TUBER CROPS Leek Lesser yam Onion True Potato Seed TP;BP S TP;BP TP TP TP; BP; S 20-15 30 20-15 20-30 20-30 20-30 10 6 6 14 21 21 14 8 28 Prechill Prechill-5C 3 day light Prechill GA3 500 ppm, 24 hrs;light Light

GREELEAFYVEGETABLES Amaranth Amara1lthus spp. Asparagus Asparagus officina/is

2 Celery Coriander Lettuce Methi Parsley Parsnip Spinach

0)

3 TI' TI':I3I' TP: 131' TI'; HI' 131' 8P;TI'; S TP;Bl' TI';BP

4 20-30 20-30:20 20 20-30;20 20-30 20-30 is-tO 20-30,15-25

5 10 7 4 5 11

6 21 21 7 14 28 28 Prechill

ApiU/11gral'colclIs Coriallllrum Satil'tI/l1

LacfUcasatil'lI Trigolle!lafoclIumgraccufn
Pc/roseli lIum

crispum Pasti/laca satim Spillacia oleracea Betll I'ulgaris

7 4

Prechill 5C Prechill Prewash (multigerm, 2 hrs; genetic monogerm, 4 hrs.)

21 14

CD

Spinachheet
ROOT CROPS Carrot Celeriac Garden beet Garden rhubarb Globe artichoke Radish Turnip

Daucus carow Apium gi'al'eolclIs Beta I'lilgaris Rheum rlzaphonticum CY/larascolymus . RaplulIlussativus

TP; 131' 1'1' TP; BP; S TP 81'; S TP;BI' TI'

20-30; 20 20-30 20-30: 15-25 20-30 20-30 20-30: 20 22-30; 20

7 10 4

14 21 14 21 21

Prechill, KNO3 Prewash multigerm 7 hrs monogerm 4 hrs.

4 5

10 7

I'rechill Prechill, KNO3

Brassicarapa

2
LEGUME VEGETABLES

:)

()

Broad Ol:an Field bl:an

Li Illa bl:a n

Viciafaha f)olichos laMah

I'l1a.\'colus IlIl/alll,I' I'hllscolus c(}ccil/cu.\'

BI';S BI': S BI':S BI';S TI' 1'1'

20 20-.30: 25 20-30: 25
20-30; 20

4 4
:)

14 10 9
9

Pnxhill

Sl:arkt b~un COLE C!HWS Cuboagl:. Knol-kohl

Cauli !lower,
0 j

Hmssica olcracca n. ole/'{/('eavar. holrylisal/d var, ilalic{/ B. peldl/el/sis

a/ld cl1i/le/lsis

20-30: 20 20-'30; 20

:)

10

I'rcl:hill, KNO3

10

Prechill, KNO3

BroCl:oli

Chinl:sc cabbagc

TP

20-30; 20

:)

Prechill

Note: (1)

Prechilling : The replicates for germination are placed in contact with the moist substratum and kepi at low temperature (between 5 and laC) for upto seven days for all agricultural and vegetable seeds.
PostasSi1l11l nitrate (KNO3) ;Instead of water 0.2% KNO3 solution (prepared by dissolving 2g KNO3 in one litre of water) is used to saturate the germination substratum at the beginning of the test. Water is used for moistening thereafter,

(2)

(3)

Gibberellic acid (GA.~): Required concentration should be prepared. For preparing 1000 ppm solution dissolve 1

gm GA3 in 1000 ml ofl120, for 500 ppm dissolve 500 mg in 1000 ml of water and for 100 ppm, 100 mg should be dissolved in 1000 ml of water. When concentration of GA3 is not mentioned, any concentration ranging from tOO to 500 rrm should be used. Seeds should be soaked in required concentration of GA,for dried on the laboratory table and rut for " germination. 17 hrs at room temperature,

">I ....

5.1

MONOCOTYLEDONS

Triticum
hYPOge~/ge~~i~~~o~

lea

Allium
epi.geo.l g,,~ in.o.cioll.

9
r.4.;' L-u .
-..j I\)

s
e 12 II I.
"11 ~

j=r,

" t...

,.,.- .

~
"::."

r-4

'/--

~
"

;:,:.

7777TT7T1,

nr

77"17"?7

rr
10

....

I.SEED
I.ADVENTITfOUS 2.COLEOPrILE 3.COTYLEDON 4.EMBRYO

WITH
RDOTS

EMBRYO
5.ENDOSPERM 6.LATERAL 8.PLUMULE ROOT 7.MESOCOTYLE

III DEVELOPED SEEDLING

PRIMARY LEAF

'3.SEED

COAT

10. PRIMA~V ROOT II. RADICLE 11'SCU.~ELLUM

14.SEMINAL ROOT

I{".#l

....

5.2
Pisum

DICOTYLEDONS Helianthus
ep"3ecd

Phaseolus
gert><L>1.t1riOI1.

fG9

I~

f~

L(B}
II 10

I- .. 5 :.:.

I_" I

."
4

"

r. J\

~':.;'

@J='"
IZ

10

h~Po9"",1ger",i"a.t:iOI1.

C') !'-

11 11
, I.EMBRYO 1 ADVENTITIOUS "ZAXILLARY 3 AXILLARY A COTYLEDON fc~ BUD SHOOT ROOTS
II .YOUN(',

SEEDLIN(',

III.DEYELOPED & LATERAL 9 PLUMULE 10 PRIMARY

SEEDLlN(', ROOT LEAF 11 PRrMARY ROOT IZRADICLE 13 SHOt'JT APEX

5 EM BRYO & E.PI COTY L 7 HYPOCOTYL

~'

5.3
SEEDLINGS OF ORYZA SATlV"

J
G

~
A. NORMAL SEEDLING: PLUMULE AND ROOTS WELL
PLUMULE AND

DEVELOPED

B. ABNORMAL SEEDLING: WEAK

RooTS

C. ABNORMALSEEDLING:
D.ABNORMAL E.ABNORMAL SEEDLING: SEEDLING:

NO
ONLY WEAK

ROOTS ONE ROoT PLUMULE THAIN ONE..HALF LENGTH OF COLEoPTlLEi

AND ROOT DEVE LOPME NT

F.ABNORMALSEEDLINb: PLUMULE LESS

G.ABNORMAL SEEDLING: WEAK PLUMULE

74

.......

S".-t 5.4

SEEDLING

OF

TRITICUM

SPP.

(7\
E F G

/~
A. B. NORMAL SEEDLI NG NORMAL SEEDLING SEEDLI NG ; GOOD ROOT AND TWO ROOTS WELL PLUMUL E DEVELOPMENT ROOTS

. .

DEVELOPED AND

SEM INAL SHORTENED

C. ABNORMAL

THICKENED

D. ABNORMAL SEEDLING E. F. ABNORMAL SEEDLING ABNORMAL SEEDLING

: EMPTY

COlEOPTIlE
THAN ONE-HALF LENGTH OF COLEOPrI-lE

: PLUMULE LESS

: WEAK : WEAK

PLUMULE
ROOTS

G. ABNORMAL SEEDLING

75

5.5

s:-SEEDLINGS OF SORGHUM Spp.

t
A. NORMALSEEDLING :WELL DEV ELOPED
B.ABNORMAL SEEDLING; WEAK EMPTY ROOTS AND PLUMULE

ROOTS
COLEOPTILE

C.ABNORMAJ. SEEDLING: D. ABNORMAL SEEDLING' E.ABNORMAL SEEDLING:

F .ABNORMAL SEEDLING:

NO ROOTS NO c.OLEOPTILE
PWMULE LESS THAN

AND PLUMULE
ONE-HALF LENGTH OF COLEOPTILE

76

5.6

SEEDLINGS

OF

MAyS

A- NCRMAL SEED\:.ING . GOOD PLUMULE '8. AII"NORMALSEEDLING' C. ABNORMAL SEEDLING: D.ABNORMAL SEEDLING: EMPTY WEAK

AND ROOT

DEVELOPMEN

COLEOPTI LE ROOT DEVELOPMENT PLUMULE

SMALl,WEAK

77

5.7
SEEDLINGS OF PHASEOLUS VULc;.ARI S D

C A B

f7)

A
B C

- NORMAL SEEDLING: EPICOTYL HYPOCorYL AND - ABNORMAL SEEDLING :BALD-HEAD i NO TERMINAL

- ABNORMAL
SEEDLING:TERMINAL BUO PRESENT

RADICLE

WELL DEVE LOPED

SUDiNO PRII4ARY LEAVES

BUT NO PRIMARY j STUNTED LEAVES

D-

ABNORMAL SEEDLIHG:PRIMARY

LEAVES

CHLOROTIC

E- NORMAL F- ABNORMAL
G

SEEDLING: ONE PRIMARY LEAF SEEDLINc.: HYPOCOTyL

PRESENT

THICKENED i NO RADICLE WEAK; NO

RADIC LE

- A BN ORMAL SEEDLI

HG : tly POCOf'(L

78

5.8
SEED LING S OF PISUM SATIVUM

RADICLE

~
SEEDLING: GOOD EPICOTYL STU NTE D AND RADICLE DEVELOPMENT SEEDLlNe.: SEEDLING: SEEDLING: SEEDLING: SEEDLING: RADICLE DEVELOPMENT EPICOTYL AND RADICLE NO RAD1tLE SPLIT/SWOLLEN WEAK NO EPICOTYL EPICOTYL

.&.JB_NORMAL C

...ABNORMAL -ASNORMAL

D
E F G

ABNORMAL

-ABNORMAL _ABNORMAL

79

5.9
SEEDLINGS OF ARACHIS HYPOGAEA

~,
A. NORMALSEEDUNG :EPICOTYL,HYPOCOTYL 8. A8NORMALSEEDLING:INSUFFICIENT AND RADICLE WELL DEVELOPED DEVELOPMENT OF SPLIT HYPOCOTYL
COTYLEDONS

PRIMARY

ROOT

C. ABNORMAL SEEDLING :THICKENED

D. ABNORMALSEEDLING: SHORT THt1:KENED HYPOCOTYL; UNOPaJED E .ABNORMAL SEEDLING: SHORT

F. ABNORMAL G .ABNoRMAL

THICKENED

HYPOCOTYL

SEEDLING: NO

RADICLE RADICLE

SEEDLING: LOOPED HYPOCOTYL; WEAK

80

/'

5.10

SEEDLINGS

Of"

HELIANTHUS

ANNUUS

r.
A

$'
E

~'
A. NORMAL SEEDLING: EPICOTyl tJYPOCOTYL AND RADICLE WELL DEVELOPED B.A BNORMAl SEEDLlN6: PRI MARY ROOT S TURTED IN SPI TE OF SECONDARy ROOTS

C.ABNORMAL SEEDLlNG:HPOCOTYl

CONSTRICTED;

NO PRIMARY ROOT

D.ABNORMAL SEEDLING :HYPOCOTYL SHORT E.ABNORMAL SEEDLiNG., PRIMARY ROOT

F. ABNORM AL SE EDLING: HYPOCOTY L

AND tHICK;

NO PRIMARY

ROOT

DECAVED; HYPOCOTYL SHORT AND THICK.!

FORMING A WEAK SPIRAL

G.ABNORMAl SEEDLING: SPLIT

HYPOCOTYL

PRIMARY

ROOT

81

5.11

7J
[

~"
E

A. NORMAL SEEDLlN6o: GOOD SEEDLlN6o DEVELOPMENT

B. NORMAL SEEDLING: ONE COTYLEDON DEV E LOP M ENT OTHERWISE &OOD SEEDLI NG

C. ABNORMAL SEEDLING: GROWIN&

D.ABNORMAL E.ABNORMAL F.ABNORMAL G.ABNORMAL H.ABNORMAL SEEDLING: SEEDLING SEEDLING: SEEDUN&: SEEDLING: SWOllEN

POINT

ABSENT

HYPOCOTYL; AREA OF

NO

RADICLE DECAYED

:SO./.OF
SPLIT THICKENED WEAK

COTYLEDONS

HYPOc.OTYl HYPOCOTYL;WEAK RADICLE

RADICLE Qii'

82

5.12
SEEDLINGS OF GOSSYPIUM SPp.

~
D

~i('
THE

A. NORMAL SEEDLlNG(SEEDCOAT OFF) :HYPOCOTYL,RADICLE AND COTYLEDONS WELL DEVELOPED B. NORMAL SEEDLlNG( SEEDCOAT ON ): SAME AS C. ABNORMAL SEEDUNG D. ABNORMAL SEEDLING ABOVE

'. RADICLE FAILED TO DEVELOP

HYPOCOTYL NOT EL OHGATED RADICLE MISSING

Eo ABNORMAL F. ABNORMAL

SEEDLING SEEDUNG

:HYPOCOTYL THICKENED j RADICLE WEAK

: HYPO COTYL DECAYED DECAYED i BASE OF COTYLEDONS

#'

83

"

5.13
OF ALLIUM

SEEDLI

N GS

SPP.

//

liB

-.;

l'

l'

7'
A. NORMAL SEEDLING! SEED COATON): COTyLEDON "eENT~ LONG;
e.. NORMAL SEEDLINGISEEDCOAT ; NO
ROOT WELL DEVELOPED

OFF): COTYLEDON'BENT'LONGjROOT

WELL

DEVELOPED

C. ABNORMAL SEE~LlNG D. ABNORMAL SEEDLING: E.ABNORMAL SEEDLING:

RADICLE SHORT, WEAK "KNEE'

COTYLEDON INDEFINITE

F. ABNORMALSEEDLING: WEAK RADICLE

84

5.14
SEEDLINGS OF

",

CITRULLUS If,

SPP.

II

,~

()
B

A.

NORMAL SEEDUNG(SEEDCOAT ON)', GOOD SEEDLING DEVELOPMENT

B. NORMAL SEEDLING! SEEDCOAT OFF): GOOD SEEDLING DEVELO,.MENT C. ABNORMALSEEDLlN6:WEAK RADICLE 'DEVELOPMENT D.ABNORMAL SEEDLING: THICKENED E. ABNORMAL SEEDLING:
F. ABNORMAL SEEDLING

HYPOCOTYL
HYPOCOT YL

SHORTENED

: COTYLEDON AND EPICOTYL ABSENT

85

/
~

86

87

88

I
I

89

"""'--

90

91

1 I

92