ESTABLISHMENT OF KURA AND RED CLOVER ON SOIL AND MINE SPOIL David C. Ditsch, Michael Collins and Norman L. Taylor
     Numerous benefits result from the addition of forage legumes to 
livestock diets.  Superior nutritive value and intake of legume forage 
generally increase individual animal productivity compared with grass 
alone. This is particularly important with tall fescue where dilution 
of the diet with legumes can partially alleviate toxicosis associated 
with endophyte fungus infected fescue (Acremonium coenophialum).  
Unfortunately, short-term persistence of most legumes requires periodic 
reestablishment, adding to the cost of forage production and complicating
pasture management.
     Perennial legumes are beneficial to soil development and the 
revegetation of reclaimed surface mined lands (Ditsch and Collins, 1996).  
After surface mining, there is little or no organic matter in the soil and 
symbiotic nitrogen (N2) fixation by legumes can provide the N to initiate 
the N cycle.  Persistent legumes could increase the overall level of plant 
available soil N during reclamation as well as improving forage yield and 
quality where the designated post-mine land use is pasture/hayland or 
wildlife habitat.
     Kura clover (Trifolium ambiguum Bieb.) is a long-lived perennial 
legume introduced from the Caucasian Region of Russia.  Kura clover 
closely resembles white clover but spreads vegetatively by rhizomes 
(horizontal underground stems).  Kura's persistence is due primarily 
to it's ability to spread over substantial distances by way of rhizome 
growth.  Researchers have reported kura clover survival after 5 years 
to be greater than white clover, red clover and alsike clover 
(Trifolium hybridum L.)  In 9-yr-old stands, kura clover plants had 
spread an average of 23 inches, with the greatest spreading occurring 
in pastures subjected to higher stocking rates (Allan and Keoghan, 1994).
     Low seedling vigor has been identified as a limitation to successful 
establishment of kura clover (Taylor and Henry, 1989; Caradus, 1994; Ehlke 
et al., 1994).  Failure to effectively inoculate kura clover seed or use of 
less effective strains of rhizobia may contribute to the observed low 
seedling vigor of kura clover (Strachan et al., 1994).  When inoculant 
rate was increased to five times the recommended rate, the percentage of 
kura clover seedlings nodulated increased from less than 10% to more than 
60%.  Seedling density increased from less than 0.5 plants/ft2 to more than 
2.8 plants/ft2.
     Objectives of this study were to compare seedling growth and stand 
development characteristics of kura clover with those of red clover on 
soil and mine spoil and to assess the potential for improvement of kura 
clover establishment by commercial seed treatments.

                      MATERIALS AND METHODS

     Plot studies were established on a Maury silt loam (Typic hapludult) 
soil on the Spindletop Research Farm near Lexington, KY and on a coal mine 
spoil site in the Appalachian coal fields of eastern Kentucky, near 
Quicksand.  The Maury soil had a pH of 6.2, > 200 lbs/acre available P 
and 281 lbs/acre of exchangable K and received no fertilization during 
the term of this experiment. Mine spoil material was a mixture of sandstone 
and shale with a pH of 7.9 and Mehlich III extractable P level of 7 
lbs/acre and more than 500 lbs/acre of exchangeable K.  This site received
158 lbs/acre P2O5 at the time of seeding.
      Treatments consisted of Kura clover seed uncoated, coated, or 
coated plus fungicide and red clover.  Seed of  Rhizo' kura clover was 
obtained from the U.S.D.A. Natural Resource and Conservation Service, 
Plant Materials Center in Quicksand, KY.  Uncoated kura clover (K) was 
inoculated with the appropriate strain of Rhizobium (LiphaTech Inc., 
Milwakee, WI) and refrigerated for 3 days prior to seeding. Portions of 
the same seed lot were coated with a commerial seed treatment (Rhozo-Kote, 
Celpril, Manteca, CA) (KC) and the same coating material plus a fungicide, 
metalaxyl (N-(2,6-Dimethylphenyl)-N-(methoxyacetyl)-alanine methyl ester 
(KCF).  Both coating treatments included the peat based Rhizobium inoculum 
at a rate of 0.4% w/w.  Germination averaged 79, 75 and 76% for uncoated, 
coated and coated kura clover plus fungicide, respectively, with not more 
than 5% hard seed.  Kenstar' red clover had a germination of 95%.
     Plots measured 2.6 x 18 ft on the Maury soil site and 5 x 20 ft on 
the mine site.  Each treatment was replicated 6 times on the Maury soil and 
4 times on the mine site, in a randomized complete block design.  Uncoated 
kura clover (K) was seeded at a rate of 12 lbs/acre using a small plot 
drill with a 6 inch row spacing.  Seeding rates of KC and KCF seed were 
increased 1.34 and 1.44 times that of uncoated seed to account for the 
weight of coating materials.
      Mine spoil plots were seeded on 29 April 1994 into a seedbed 
prepared by using a rotary tine tiller.  An earlier seeding on 19 April 
1994 on the Maury soil failed due to a series of heavy rain events during 
a 2 week period after seeding, and a second seeding on the Maury soil was 
made on 24 May 1994.  No herbicides were used on the mine site.  Methyl 
bromide (bromomethane) was used over the Maury soil plot area.  Red clover 
shoot growth was removed twice during 1994 by clipping at a 2.5 inch 
stubble height.  Kura clover plots were also clipped but little biomass 
was removed.  Shoot growth was left unclipped on the mine spoil plots in 
accordance with typical management on such sites.
     Seedling density was determined by counting the number of live 
seedlings in two randomly selected 1 meter (39 inches) lengths of row in 
each plot.  Shoot mass was determined by cutting 10-15 consecutive plants 
at the soil surface from a randomly selected starting point in each plot.  
Final stand density determinations and seedling mass samples were collected 
on 4 and 5 May 1995 from the Maury soil and mine spoil sites, respectively. 
At the same time, ten whole plants were collected from each plot by digging 
to a depth of 12 inches. After washing, each plant was separated into 
below-ground components, which included the root and rhizomes, crown and 
shoot components.  Each sample was dried at 140o F for 72 hr before 
weighing.  Daughter plants (new plants initiated by the original seedling) 
were counted, removed, separated into shoot, crown and root components, and 
dried as described above.  Grazing damage by wildlife precluded sampling on 
the mine spoil site.
     Location effects were considered fixed and sampling date effects 
were analyzed as repeated measures (SAS, 1985).  Contrasts and multiple 
comparison test were used to compare coated and uncoated kura clover 
(K vs. KC) and to compare red clover and kura clover.

                      RESULTS AND DISCUSSION

Weather Conditions

     Precipitation during 1994 was generally below long-term mean levels 
at both locations, with deficits during 6 of the 8 months between May and 
December at both locations.  July precipitation was 2.4 and 1.6 inches 
below the long-term mean on the soil and mine sites, respecitively.  Mean 
monthly air temperatures during fall and winter following seeding at the 
mine site were 3.1 to 10.1o F above normal after being near normal through 
September.  Mean monthly temperatures averaged 3.1o F below normal during 
Aug. and Sept. on the soil site but were also generally above normal otherwise.

Stand Density

      Red clover consistently had greater stand densities than kura clover 
at both location. (Fig. 1).  Maximum red clover stand densities near 18 
plants per ft of row at both sites occured within the first and second 
sampling dates.  Maximum kura clover stand densities also occured within 
the first two sampling dates (2-3 weeks after planting) and ranged between 
8 and 11 plants per ft of row at the Maury soil and mine site, respectively.
The stand density of 18 red clover seedling per ft of row represents 
emergence of slightly more the one-half (53%) of the live seeds, based on 
a 12 lbs/ac seeding rate and 95% germination.  At 77% germination and the 
same seeding rate, a kura clover stand density of 11 plants per ft of row 
represents only 33% of the live seeds.  This difference suggest that low 
emergence may contribute to delayed or inadequate establishment of kura 
clover compared with red clover.
      By August of the seeding year, red clover stand densities on the 
Maury soil site had declined sharply to about one-third of peak densities,
compared to only a 26% decline on the mine site.  Differences in stand 
reduction between the two sites is likely due to greater competition 
imposed by the much larger red clover seedlings present on the Maury 
soil site.
     Except for greater seedling stand densities recorded for KCF than 
for K in August of the seeding year on the mine site, seed coating had 
no effect on kura clover stand density (Fig. 1). Across coating treatments,
an average of 2.8 kura clover seedlings per ft of row were present at the 
mine site on the last sampling date in the year of seeding.  Measured 1 yr 
after seeding on mine spoil, uncoated kura clover averaged 1.2 plants per 
ft of row compared to 6.2 plants per ft of row for red clover (Fig. 1).  
After the same time period on the Maury soil site, uncoated kura clover 
had 3.1 plants per ft of row compared with 8.3 plants per ft of row for 
red clover.

Yield Components

     Red clover shoot mass, measured in June and August of the seeding 
year, was significantly greater than that of kura clover at both 
locations (Table 1).  In June, red clover on the Maury soil site 
averaged 13.6 times more shoot mass that kura clover.  On the same 
date, red clover on the mine site had 4.1 times more shoot mass per 
seedling than kura clover.  Due at least in part to summer moisture 
stress on the mine site, kura clover shoot mass increased from 6.6 mg 
per plant in June to 10.9 mg per plant in August.  During approximately 
the same interval at the Maury soil site, kura clover shoot mass 
increased from 8.9 to 222 mg per plant.  Seed coating treatment had 
no effect on kura clover seedling mass measured the year of seeding.
     One year after seeding, kura clover treatments on the Maury soil 
site had greater total-seedling, crown, shoot and root/rhizome masses 
than those on the mine site (Fig. 2).  Although red clover from the Maury 
soil site also tended to have greater component and total plant masses 
than that from the mine site, the difference was much less than for kura 
clover.  One-year-old red clover seedlings on the Maury soil site were 
1.8 times heavier than those from mine spoil, but kura clover plants were 
nearly 16 times heavier on the soil site.
     Slow stand development may also be related to the tendency of kura 
clover to partition photosynthate preferentially to root and rhizome 
tissues compared with other legumes (Strachan et al., 1994).  We found 
that 1-year-old kura clover on the mine site contained 49 to 58% of its 
total mass below ground compared with only 28% below ground for red 
clover from the same site.  Both species had lower proportions of their 
total mass below ground on the Maury soil site than on the mine site 
(Table 2).  
     Two-year-old kura clover parent plants on the Maury soil site 
averaged 2.4, 1.1 and 2.9 g per plant for shoot, crown and root tissue, 
respectively (Fig. 3).  No coating or fungicide effects were found for 
kura clover plant component weights measured 2-yr after seeding.  Red 
clover was omitted from this sampling due to thin stands.  On average, 
8.9 daughter plants had been initiated on each parent plant, each with 
an average of 0.37 g of shoot mass.  The small mass of root tissue found 
on daughter plants suggests that the parent plants may still play an 
important role in daughter plant survival.

Rhizome Number and Size

     Moorhead et al. (1994) measured rapid kura clover rhizome growth 
on an acid, low-P soil using a strip tillage planting method.  Within 
5 months after seeding, 75% of kura clover plants had rhizomes averaging 
0.82 inches in length. In the our study, two-year-old kura clover plants 
had an average of 12.5 rhizomes per plant with average lengths of 6 inches 
per rhizome (Fig. 3).

Conclusion

     Kura clover dry matter production during the seeding year was much 
less than red clover at both locations. However, red clover seedling 
density declined more rapidly than kura clover at both locations during 
this same period suggesting greater kura clover persistence.  These data 
also indicate that low kura clover dry matter yields, compared to red 
clover, may be related to greater below ground growth, particularly on 
mine spoil where frequent periods of moisture stress are common.  In 
general, seed coatings had no effect on growth and persistence of kura 
clover at both locations.
     Greater kura clover persistence along with the potential for 
higher dry matter production over time, compared to red clover, are 
encouraging results, particularly for use on reclaimed surface mined 
land.  However, more research is needed to identify management strategies 
that increase the rate of establishment and seed mixtures that are 
compatible to the slow development of kura clover. 



Table 1.  Shoot dry weight of kura and red clover seedlings sampled 
during the year of seeding.

_________________________________________________________________
   Location        Seed coating/species        June     August
_________________________________________________________________
                                              ....mg/seedling....

Mine Spoil    Kura (K)                         8.0 b     9.2 b

              Kura-coated (KC)                 6.7 b    13.8 b

              Kura-coated + fungicide (KCF)    5.2 b     9.7 b

              Red clover (RC)                 27.1 a    83.2 a




Maury silt loam soil  Kura                    12.7 b   249 b

              Kura-coated                      5.6 b   143 b

              Kura-coated + fungicide          8.5 b   273 b

              Red clover                     121.3 a  1901 a
_________________________________________________________________

Means within a column and location followed by the same letter are
 not significantly different at  the 0.05 level based on the LSD test.



Table 2.  Plant component yields of kura and red clover one year 
after seeding.
_________________________________________________________________
                                       Plant Component
                                _________________________________
Location      Treatment          Shoot   Crown   Root     Total
_________________________________________________________________

                                    ....% of Total.....    lbs/ac

Mine Spoil   Kura (K)           26.3 b  15.9 b  57.8 b    107.4 b

          Kura-coated (KC)      30.3 b  13.6 b  56.2 b    130.4 b

  Kura-coated + fungicide (KCF) 34.7 b  16.1 b  49.2 b    137.5 b

          Red clover (RC)       45.4 a  26.4 a  28.3 a   2368.2 a



Maury silt    Kura              42.2 b  22.2 a  35.5 a   3805.9 b
loam soil
              Kura-coated       42.5 c  21.3 b  36.2 bc  2008.4 c

  Kura-coated + fungicide (KCF) 44.3 c  21.1 b  34.6 c   1914.6 c

          Red clover (RC)       56.4 a  20.6 a  23.0 ab  5499.1 a
_________________________________________________________________
 Means within a column followed by the same letter are not 
significantly different at the 0.05 level based on the LSD test.
  Means within a column and location, followed by the same letter, 
are not significantly different at the 0.05 level based on the LSD test.



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Caradus, J.R. 1994. Frost tolerance of Trifolium species. New Zealand 
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Ditsch, D.C. and M. Collins. 1996. Reclamation considerations for 
pasture and haylands receiving 66 cm or more precipitation annually.
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Strachan, D.E., A.H. Nordmeyer, and J.G.H. White. 1994. Nutrient 
storage in roots and rhizomes of hexaploid caucasian clover.  Proc. 
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Taylor, N.L., and D. Henry. 1989. Kura clover for Kentucky.  University 
of Kentucky Agric. Exp. Stn. Pub. AGR-141.